Secreted factors

ABSTRACT

The invention concerns new secreted factors encoded by clones P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_CO1 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), P00697_C03 (SEQ ID NO:75), and other mammalian homologues and variants of such factor, as well as polynucleotides encoding them. The invention further concerns methods and means for producing such factors and their use in the diagnosis and treatment of various cardiac, renal or inflammatory diseases.

SECRETED FACTORS

[0001] This application claims benefit under Title 35, United StatesCode §119(e) of U.S. provisional application No. 60/193,548 filed onMar. 31, 2000.

FIELD OF THE INVENTION

[0002] The present invention concerns secreted factors encoded by genesdifferentially regulated in certain diseased tissues. More particularly,the invention concerns nucleic acid encoding novel secreted polypeptidefactors, the encoded polypeptides, and compositions containing andmethods and means for producing them. The invention further concernsmethods based on the use of such nucleic acids and/or polypeptides inthe diagnosis and treatment of various diseases, in particular cardiac,renal, or inflammatory diseases.

BACKGROUND OF THE INVENTION

[0003] Gene expression patterns, including changes in gene expressionbetween normal and diseased tissues or tissues in various stages ofdisease progression provide valuable insight into the moleculardeterminants of normal and abnormal cellular physiology. Accordingly,genes that are differentially expressed in subjects suffering from adisease, such as cardiac, renal or inflammatory disease, relative tonormal subjects, are useful targets for intervention to diagnose,prevent or treat such diseases.

[0004] Techniques have been developed to efficiently analyze the levelof expression of specific genes in cells and tissues. Procedures thatcan be used to identify and clone differentially expressed genesinclude, for example, subtractive hybridization (Jiang and Fisher, Mol.Cell. Different. 1:285-299 [1993]; Jiang etal., Oncogene 10, 1855-1864[1995]; Sagerstrom et al., Annu. Rev. Biochem. 66: 751-783 [1997]);differential RNA display (DDRT-PCR) (Watson et al., DevelopmentalNeuroscience 15:77-86 [1993]; Liang and Pardee, Science 257:967-971[1992]); RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) (Ralphet al., Proc. Natl. Acad. Sci. USA 90:10710-10714 [1993]; McClelland andWelsh, PCR Methods and Applications 4:S66-81 [1994]); representationaldifference analysis (RDA) (Hubank and Schatz, Nucl. Acids Res.22:5640-5648 [1994]); serial analysis of gene expression (SAGE)(Velculescu et al., Science 270:484-487 [1995]; Zhang et al., Science276:1268-1272 [1997]); electronic subtraction (Wan et al., NatureBiotechnologyl4:1685-1691 [1996]); combinatorial gene matrix analyses(Schena et al., Science 270:467-470 [1995]), and various modificationsand improvements of these and similar techniques.

[0005] A particularly attractive method for assessing gene expression isthe DNA microarray technique. In this method, nucleotide sequences ofinterest are plated, or arrayed, on a porous or non-porous substratethat can be paper, nylon or other type of membrane, filter, chip, glassslide or any other suitable solid support. The arrayed sequences arethen hybridized with specific DNA probes from cells or tissues ofinterest. Microarrays of biological materials have been described in anumber of patents and patent applications, including, for example, U.S.Pat. Nos. 5,744,305; 5,800,992; 5,807,522; 5,716,785; and EuropeanPatent No. 0 373 203.

[0006] The DNA microarray technique can be used to monitor theexpression level of large numbers of genes simultaneously (to produce atranscript image), and to identify genetic variants, mutations andpolymorphisms. This information may be used to determine gene function,understanding the genetic basis of disease, diagnosing disease, anddeveloping and monitoring the activities of therapeutic agents.

[0007] An important application of the microarray method allows for theassessment of differential gene expression in pairs of mRNA samples fromtwo different tissues, or in the same tissue comparing normal versusdisease states or time progression of the disease. Microarray analysisallows one to analyze the expression of known genes of interest, or todiscover novel genes expressed differentially in tissues of interest.Thus, an attractive application of this technology is as a fundamentaldiscovery tool to identify new genes, and their corresponding expressionproducts, which contribute to the pathogenesis of disease and relatedconditions.

[0008] Microarray technology has been successfully applied tolarge-scale analysis of human gene expression to identifycancer-specific genes and inflammatory-specific genes (DeRisi et aL,Nat. Genet., 14(4):457-60 [1996]; Heller et al., Proc. Natl. Acad. Sci.USA, 94(6):2150-55 [1997]). DeRisi et al. examined a pre-selected set of870 different genes for their expression in a melanoma cell line and anon-tumorigenic version of the same cell line. The microarray analysisrevealed a decrease in expression for 15/870 (1.7%) and an increase inexpression for 63/870 (7.3%) of the genes in non-tumorigenic relative totumorigenic cells (differential expression values <0.52 or >2.4 weredeemed significant). Heller et al. employed microarrays to evaluate theexpression of 1000 genes in cells taken from normal and inflamed humantissues. The results indicated that altered expression was evident ingenes encoding inflammatory mediators such as IL-3, and a tissuemetalloprotease. These results illustrate the utility of applyingmicroarray technology to complex human diseases.

[0009] It would be beneficial to discover differentially expressed genesthat are related to diseases or various disease states. It would furtherbe beneficial to develop methods and compositions for the diagnosticevaluation and prognosis of conditions involving such diseases, for theidentification of subjects exhibiting a predisposition to suchconditions, for modulating the effect of these differentially expressedgenes and their expression products, for monitoring patients undergoingclinical evaluation for the prevention and treatment of a disease,specifically cardiac, kidney or inflammatory disease, and for monitoringthe efficacy of compounds used in clinical trials.

[0010] Secreted proteins mediate key biological processes including cellto cell interactions as well as important cellular functions such ascell growth and differentiation, and most protein-based drugs aresecreted proteins including insulin, growth hormone, interferons, tissueplasminogen activator ( tPA), and erythropoietin (EPO). It would,therefore, be particularly desirable to identify novel differentiallyexpressed genes encoding secreted proteins.

SUMMARY OF TIE INVENTION

[0011] In one aspect, the present invention concerns an isolated nucleicacid molecule comprising a poly-or oligonucleotide selected from thegroup consisting of:

[0012] (a) a polynucleotide encoding a polypeptide having at least about80% sequence identity with amino acids selected from the groupconsisting of: 1 to 1203 of SEQ ID NO:2, amino acids 1 to 193 of SEQ IDNO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 ofSEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to82 of SEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1to 170 of SEQ ID NO:24, amino acids −26 to 233 of FIG. 13 (amino acids 1to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, aminoacids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO: 33,amino acids 1 to 30 of SEQ ID NO:35, amino acids 1 to 100 of SEQ IDNO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 ofSEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids −35 to387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, aminoacids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69,amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1 to 114 of SEQID NO:74, and amino acids 1 to 97 of SEQ ID NO:76; or a transmembranedomain (membrane spanning segment/region) deleted or inactivated variantthereof;

[0013] (b) a polynucleotide encoding a polypeptide having at least about80% sequence identity with amino acids 1 to 233 of SEQ ID NO: 26, oramino acids 1 to 387 of SEQ ID NO: 53;

[0014] (c) a polynucleotide encoding amino acids selected from the groupconsisting of: 1 to 203 of SEQ ID NO: 2, amino acids 1 to 193 of SEQ IDNO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 ofSEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to82 of SEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1to 170 of SEQ ID NO:24, amino acids −26 to 233 of FIG. 13 (amino acids 1to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, aminoacids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO: 33,amino acids 1 to 30 of SEQ ID NO:35, amino acids 1 to 100 of SEQ IDNO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 ofSEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids −35 to387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, aminoacids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69,amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1 to 114 of SEQID NO:74, and amino acids 1 to 97 of SEQ ID NO:76; or a transmembranedomain (membrane spanning segment/region) deleted or inactivated variantthereof;

[0015] (d) a polynucleotide selected from the group consisting of: apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 1, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity o [the polypeptide encoded by clone P00184_D11 (SEQ ID NO: 1),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 3, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00185_D11 (SEQ ID NO: 3);a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 5, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone POO1 88D12 (SEQ ID NO: 5),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 7, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 9, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_G01 (SEQ ID NO: 9),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 11, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_G05 (SEQ ID NO: 11),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 13, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_H10 (SEQ ID NO:13),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 15, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone POO199_D08 (SEQ ID NO: 15),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 17, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00203_D04 (SEQ ID NO: 17),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 19, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00203_E06 (SEQ ID NO: 19),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 21, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00209_F06 (SEQ ID NO: 21),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 23, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00219_D02 (SEQ ID NO: 23),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 25, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 27, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00220_H05 (SEQ ID NO: 27),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 29, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00222_G03 (SEQ ID NO: 29),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 31 (clone P00223_F07), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 32, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00225_C01 (SEQ ID NO: 32),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 34, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00227_D11 (SEQ ID NO: 34),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 36, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00228_F03 (SEQ ID NO: 36),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 38, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00233_H08 (SEQ ID NO: 38),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 40, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00235_G08 (SEQ ID NO: 40),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 42, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00239_C11 (SEQ ID NO: 42),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 44 (clone P00240_B04), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 45, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00240_E05 (SEQ ID NO: 45),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 47 (clone P00241_E12), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 48 (clone P00245_D06), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 49 (clone P00246_D12), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 50, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00247_A04 (SEQ ID NO: 50),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 52, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00248_B04 (SEQ ID NO: 52),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 54, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00249_F09 (SEQ ID NO: 54),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 56, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00258_A10 (SEQ ID NO: 56),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 58, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00262_C10 (SEQ ID NO: 58),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 60 (clone P00263_G06), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 61 (clone P00267_F08) , apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 62, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00269_H08 (SEQ ID NO: 62),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 64 (clone P00312_C04), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 65 (clone P00324_H02), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 66, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00628_H02 (SEQ ID NO: 66),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 68, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00629_C08 (SEQ ID NO: 68),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 70 (clone P00634_G11), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 71, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00641_G11 (SEQ ID NO: 71),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 73, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00648_E12 (SEQ ID NO: 73),and a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 75 wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00697_C03 (SEQ ID NO: 75);

[0016] (e) a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids selected from the group consisting of: aminoacids 1 to 203 of SEQ ID NO: 2, wherein said polynucleotide encodes apolypeptide having at least one biological activity of the polypeptideencoded by clone P00184_D11 (SEQ ID NO: 1), a polynucleotide encoding atleast about 50 contiguous amino acids from amino acids 1 to 193 of SEQID NO: 4, wherein said polynucleotide encodes a polypeptide having atleast one biological activity of the polypeptide encoded by cloneP00185_D11 (SEQ ID NO: 3); a polynucleotide encoding at least about 50contiguous amino acids from amino acids 1 to 236 of SEQ ID NO: 6,wherein said polynucleotide encodes a polypeptide having at least onebiological activity of the polypeptide encoded by clone P00188_D12 (SEQID NO: 5), a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids 1 to 61 of SEQ ID NO: 8, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 79 of SEQ ID NO: 10, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00194_G01 (SEQ ID NO: 9), a polynucleotideencoding at least about 50 contiguous amino acids from amino acids 1 to92 of SEQ ID NO: 12, wherein said polynucleotide encodes a polypeptidehaving at least one biological activity of the polypeptide encoded byclone P00194_G05 (SEQ ID NO: 11), a polynucleotide encoding at leastabout 50 contiguous amino acids from amino acids 1 to 86 of SEQ ID NO:14, wherein said polynucleotide encodes a polypeptide having at leastone biological activity of the polypeptide encoded by clone P00194_H10(SEQ ID NO:13), a polynucleotide encoding at least about 50 contiguousamino acids from amino acids 1 to 36 of SEQ ID NO: 16, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00199_D08 (SEQ ID NO: 15),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 83 of SEQ ID NO: 18, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00203_D04 (SEQ ID NO: 17), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 82 of SEQ ID NO: 20, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00203_E06 (SEQ ID NO: 19), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 462 of SEQ ID NO: 22, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00209_F06 (SEQ ID NO: 21), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 170 of SEQ ID NO: 24, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00219_D02 (SEQ ID NO: 23), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids −26 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ IDNO:26), wherein said polynucleotide encodes a polypeptide having atleast one biological activity of the polypeptide encoded by cloneP00219_F06 (SEQ ID NO: 25), a polynucleotide encoding at least about 50contiguous amino acids from amino acids 1 to 30 of SEQ ID NO: 28,wherein said polynucleotide encodes a polypeptide having at least onebiological activity of the polypeptide encoded by clone P00220_H05 (SEQID NO: 27), a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids 1 to 39 of SEQ ID NO: 30, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00222_G03 (SEQ ID NO: 29),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 541 of SEQ ID NO: 33, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00225_C0l (SEQ ID NO: 32), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 30 of SEQ ID NO: 35, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00227_D 11 (SEQ ID NO: 34), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 100 of SEQ ID NO: 37, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00228_F03 (SEQ ID NO: 36), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 65 of SEQ ID NO: 39, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00233_H08 (SEQ ID NO: 38), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 65 of SEQ ID NO: 39, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00235_G08 (SEQ ID NO: 40), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 46 of SEQ ID NO: 43, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00239_C11 (SEQ ID NO: 42), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 313 of SEQ ID NO: 46, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00240_E05 (SEQ ID NO: 45), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 58 of SEQ ID NO: 51, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00247_A04 (SEQ ID NO: 50), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids −35 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ IDNO:53), wherein said polynucleotide encodes a polypeptide having atleast one biological activity of the polypeptide encoded by cloneP00248_B04 (SEQ ID NO: 52), a polynucleotide encoding at least about 50contiguous amino acids from amino acids I to 58 of SEQ ID NO: 55,wherein said polynucleotide encodes a polypeptide having at least onebiological activity of the polypeptide encoded by clone P00249_F09 (SEQID NO: 54), a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids 1 to 52 of SEQ ID NO: 57, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00258_A10 (SEQ ID NO: 56),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 245 of SEQ ID NO: 59, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00262_C10 (SEQ ID NO: 58), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 142 of SEQ ID NO: 63, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00269_H08 (SEQ ID NO: 62), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 49 of SEQ ID NO: 67, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00628_H02 (SEQ ID NO: 66), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 70 of SEQ ID NO: 69, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00629_C08 (SEQ ID NO: 68), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 113 of SEQ ID NO: 72, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00641_G11 (SEQ ID NO: 71), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 114 of SEQ ID NO: 74, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00648_E12 (SEQ ID NO: 73), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 97 of SEQ ID NO: 76, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00697_C03 (SEQ ID NO: 75);

[0017] (f) a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids 1 to 233 of SEQ ID NO: 26, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25),and a polynucleotide encoding at least about 50 contiguous amino acidsfrom amino acids 1 to 387 of SEQ ID NO: 53, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00248_B04 (SEQ ID NO: 52);

[0018] (g) a polynucleotide selected from the group consisting of SEQ IDNOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32, 34,36, 38, 40, 42, 44, 45, 47, 48, 49, 50, 52, 54, 56, 58, 60, 61, 62, 64,65, 66, 68, 70, 71, 73, and 75;

[0019] (h) the complement of a polynucleotide of (a)-(g); and

[0020] (i) an antisense oligonucleotide capable of hybridizing with, andinhibiting the translation of, the rnRNA encoded by a gene encoding apolypeptide selected from the group consisting of: SEQ ID NOS: 2, 4, 6,8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43,46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, 76, and another mammalian(e.g. human) homologue thereof.

[0021] In another aspect, the invention concerns a vector comprising anyof the poly- or oligonucleotides of (a)-(i) above.

[0022] In a further aspect, the invention concerns a recombinant hostcell transformed with nucleic acid comprising any of the poly- oroligonucleotides of (a)-(i) above, or with a vector comprising any ofthe poly- or oligonucleotides of (a)-(i) above.

[0023] In a still further aspect, the invention concerns a recombinantmethod for producing a polypeptide by culturing a recombinant host celltransformed with nucleic acid comprising any of the polynucleotides of(a)-(g) above under conditions such that the polypeptide is expressed,and isolating the polypeptide.

[0024] In a different aspect, the invention concerns a polypeptidecomprising:

[0025] (a) a polypeptide having at least about 80% identity with aminoacids 1 to 203 of SEQ ID NO: 2, amino acids 1 to 193 of SEQ ID NO: 4,amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQ ID NO:8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 of SEQ IDNO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36 of SEQID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to 82 ofSEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1 to 170of SEQ ID NO:24, amino acids −26 to 233 of FIG. 13 (amino acids 1 to 259of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, amino acids 1 to39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO:33, amino acids 1to 30 of SEQ ID NO: 35, amino acids 1 to 100 of SEQ ID NO:37, aminoacids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQ ID NO:41,amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 of SEQ IDNO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids −35 to 387 ofFIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to 58 ofSEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1 to 245of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, amino acids 1 to49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69, amino acids 1to 113 of SEQ ID NO:72, amino acids 1 to 114 of SEQ ID NO:74, aminoacids 1 to 97 of SEQ ID NO:76; or

[0026] a polypeptide encoded by nucleic acid hybridizing under stringentconditions with the complement of the coding region of SEQ ID NOS:1, 3,5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40,42, 44, 45, 47, 48, 49, 50, 52 54, 56, 58, 60, 61, 62, 64, 65, 66, 68,70, 71, 73, 75;

[0027] the polypeptides of (a) and (b) having at least one biologicalactivity of the polypeptide encoded by clones P00184_D11 (SEQ ID NO:1),P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ IDNO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10(SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17),P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ IDNO:23), P00219_F06 (SEQ ID NO:25 ), P00220_H05 (SEQ ID NO:27),P00222_G03 (SEQ ID NO:29), P00223_F07 (SEQ ID NO:31), P00225_C01 (SEQ IDNO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08(SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42),P00240_B04 (SEQ ID NO:44), P00240_E05 (SEQ ID NO:45), P00241_E12 (SEQ IDNO:47), P00245_D06 (SEQ ID NO:48), P00246_D12 (SEQ ID NO:49), P00247_A04(SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54),P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00263_G06 (SEQ IDNO:60), P00267_F08 (SEQ ID NO:61), P00269_H08 (SEQ ID NO:62), P00312_C04(SEQ ID NO:64), P00324_H02 (SEQ ID NO:65), P00628_H02 (SEQ ID NO:66),P00629_C08 (SEQ ID NO:68), P00634_G11 (SEQ ID NO:70), P00641_G11 (SEQ IDNO:71), P00648_E12 (SEQ ID NO:73), P00697_C03 (SEQ ID NO:75);

[0028] In another aspect, the invention concerns a compositioncomprising a polypeptide as hereinabove defined in admixture with apharmaceutically acceptable carrier. In a specific embodiment, thecomposition is a pharmaceutical composition, preferably for thetreatment of a cardiac, renal or inflanmmatory disease, comprising aneffective amount of a polypeptide of the present invention.

[0029] In yet another aspect, the invention concerns an antibodyspecifically binding a polypeptide of the present invention (ashereinabove defined).

[0030] In a further aspect, the invention concerns an antagonist oragonist of a polypetide of the present invention.

[0031] In a still further aspect, the invention concerns a composition,preferably a pharmaceutical composition, comprising an effective amountof an antibody herein, in admixture with a pharmaceutically acceptablecarrier.

[0032] The invention further concerns a composition, preferably apharmaceutical composition, comprising an effective amount of anantagonist or agonist of the present invention, in admixture with apharmaceutically acceptable carrier.

[0033] In a further aspect, the invention concerns a method for thetreatment of a cardiac, renal or inflammatory disease, comprisingadministering to a patient in need an effective amount of a polypeptideof the present invention or an antagonist or agonist thereof.

[0034] In a different aspect, the invention concerns a method for thetreatment of a cardiac, renal or inflammatory disease, comprisingadministering to a patient in need an effective amount of a poly- oroligonucleotide of the present invention (as hereinabove defined).

[0035] The invention also concerns a method for the treatment of acardiac, renal or inflammatory disease, comprising administering to apatient in need an effective amount of an antibody specifically bindingto a polypeptide of the present invention.

[0036] In a further aspect, the invention concerns a method forscreening a subject for a cardiac, renal or inflammatory diseasecharacterized by the differential expression of the endogenous homologueof the proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22,24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67,69, 72, 74, or 76 comprising the steps of:

[0037] measuring the expression in the subject of the endogenoushomologue of the protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59,63, 67, 69, 72, 74, or 76; and

[0038] determining the relative expression of such endogenous homologuein the subject compared to its expression in normal subjects, orcompared to its expression in the same subject at an earlier stage ofdevelopment of the cardiac, renal or inflammatory disease. The subjectis preferably human and, accordingly, the endogenous protein is a humanhomologue of the rat proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16,18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57,59, 63, 67, 69, 72, 74, or 76.

[0039] In a still further aspect, the invention concerns an arraycomprising one or more oligonucleotides complementary to reference RNAor DNA encoding a protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59,63, 67, 69, 72, 74, or 76 or another mammalian (e.g. human) homologuethereof, where the reference DNA or RNA sequences are obtained from botha biological sample from a normal subject and a biological sample from asubject exhibiting a cardiac, renal, or inflanmmatory disease, or frombiological samples taken at different stages of a cardiac, renal, orinflammatory disease.

[0040] In yet another aspect, the invention concerns a method fordetecting cardiac, kidney, or inflammatory disease in a human patientcomprising the steps of:

[0041] providing an array of oligonucleotides at known locations on asubstrate, which array comprises oligonucleotides complementary toreference DNA or RNA sequences encoding a human homologue of theproteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72,74, or 76, where the reference DNA or RNA sequences are obtained fromboth a biological sample from a normal patient and a biological samplefrom a patient potentially exhibiting cardiac, renal, or inflammatorydisease, or from a patient exhibiting cardiac, renal, or inflammatorydisease, taken at different stages of such disease (jointly referred toas “the test patient”);

[0042] exposing the array, under hybridization conditions, to a firstsample of cDNA probes constructed from mRNA obtained from a biologicalsample from a corresponding biological sample of a normal patient orfrom a test patient at a certain stage of the disease;

[0043] exposing the array, under hybridization conditions, to a secondsample of cDNA probes constructed from MRNA obtained from a biologicalsample obtained from the test patient (if the first sample was taken ata certain stage of the disease, the second sample is taken at adifferent stage of the disease);

[0044] quantifying any hybridization between the first sample of cDNAprobes and the second sample of cDNA probes with the oligonucleotideprobes on the array; and

[0045] determining the relative expression of genes encoding the humanhomologue of the protein of SEQ ID NO: 2 in the biological samples fromthe normal patient and the test patient, or in the biological samplestaken from the test patient at different stages of the disease.

[0046] The invention further concerns a diagnostic kit comprising anarray herein (as defined above) for detecting and diagnosing a disease,specifically cardiac, kidney or inflammatory disease. This kit maycomprise control oligonucleotide probes, PCR reagents and detectablelabels. In addition, this kit may comprise biological samples taken fromhuman subjects, said samples comprising blood or tissue, preferablycardiac tissue, more preferably left ventricle cells. Such diagnostickits may also comprise antibodies (including poly- and monoclonalantibodies) to a polypeptide of the present invention, including thepolypeptide of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24,26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69,72, 74, or 76 and further mammalian (e.g. human) homologues thereof.

BRIEF DESCRIPTION OF THE FIGURES

[0047]FIG. 1 shows the nucleotide sequence (SEQ ID NO: 1) of the cloneP0184_D11 and deduced amino acid sequence of the polypeptide (SEQ ID NO:2) enoded by the clone.

[0048]FIG. 2 shows the nucleotide sequence (SEQ ID NO: 3) of the cloneP0185_D11 and deduced amino acid sequence of the polypeptide (SEQ ID NO:4) enoded by the clone.

[0049]FIG. 3 shows the nucleotide sequence (SEQ ID NO: 5) of the cloneP0188_D12 and deduced amino acid sequence of the polypeptide (SEQ ID NO:6) enoded by the clone.

[0050]FIG. 4 shows the nucleotide sequence (SEQ ID NO: 7) of the cloneP0188_E01 and deduced amino acid sequence of the polypeptide (SEQ ID NO:8) enoded by the clone.

[0051]FIG. 5 shows the nucleotide sequence (SEQ ID NO: 9) of the cloneP0194_G01 and deduced amino acid sequence of the polypeptide (SEQ ID NO:10) enoded by the clone.

[0052]FIG. 6 shows the nucleotide sequence (SEQ ID NO: 11) of the cloneP0194_G05 and deduced amino acid sequence of the polypeptide (SEQ ID NO:12) enoded by the clone.

[0053]FIG. 7 shows the nucleotide sequence (SEQ ID NO: 13) of the cloneP0194_H10 and deduced amino acid sequence of the polypeptide (SEQ ID NO:14) enoded by the clone.

[0054]FIG. 8 shows the nucleotide sequence (SEQ ID NO: 15) of the cloneP0199_D08 and deduced amino acid sequence of the polypeptide (SEQ ID NO:16) enoded by the clone.

[0055]FIG. 9 shows the nucleotide sequence (SEQ ID NO: 17) of the cloneP0203_D04 and deduced amino acid sequence of the polypeptide (SEQ ID NO:18) enoded by the clone.

[0056]FIG. 10 shows the nucleotide sequence (SEQ ID NO: 19) of the cloneP0203_E06 and deduced amino acid sequence of the polypeptide (SEQ ID NO:20) enoded by the clone.

[0057]FIG. 11 shows the nucleotide sequence (SEQ ID NO: 21) of the cloneP0209_F06 and deduced amino acid sequence of the polypeptide (SEQ ID NO:22) enoded by the clone.

[0058]FIG. 12 shows the nucleotide sequence (SEQ ID NO: 23) of the cloneP0219_D02 and deduced amino acid sequence of the polypeptide (SEQ ID NO:24) enoded by the clone.

[0059]FIG. 13 shows the nucleotide sequence (SEQ ID NO: 25) of the cloneP0219_F06 and deduced amino acid sequence of the polypeptide (SEQ ID NO:26) enoded by the clone. The underlined amino acid residues at theN-terminal end represent a putative signal peptide.

[0060]FIG. 14 shows the nucleotide sequence (SEQ ID NO: 27) of the cloneP0220_H05 and deduced amino acid sequence of the polypeptide (SEQ ID NO:28) enoded by the clone.

[0061]FIG. 15 shows the nucleotide sequence (SEQ ID NO: 29) of the cloneP0222_G03 and deduced amino acid sequence of the polypeptide (SEQ ID NO:30) enoded by the clone.

[0062]FIG. 16 shows the nucleotide sequence (SEQ ID NO: 31) of the cloneP0184_D11.

[0063]FIG. 17 shows the nucleotide sequence (SEQ ID NO: 32) of the cloneP0225_C01 and deduced amino acid sequence of the polypeptide (SEQ ID NO:33) enoded by the clone.

[0064]FIG. 18 shows the nucleotide sequence (SEQ ID NO: 34) of the cloneP0227_D11 and deduced amino acid sequence of the polypeptide (SEQ ID NO:35) enoded by the clone.

[0065]FIG. 19 shows the nucleotide sequence (SEQ ID NO: 36) of the cloneP0228_F03 and deduced amino acid sequence of the polypeptide (SEQ ID NO:37) enoded by the clone.

[0066]FIG. 20 shows the nucleotide sequence (SEQ ID NO: 38) of the cloneP0233_H08 and deduced amino acid sequence of the polypeptide (SEQ ID NO:39) enoded by the clone.

[0067]FIG. 21 shows the nucleotide sequence (SEQ ID NO: 40) of the cloneP0235_G08 and deduced amino acid sequence of the polypeptide (SEQ ID NO:41) enoded by the clone.

[0068]FIG. 22 shows the nucleotide sequence (SEQ ID NO: 42) of the cloneP0239_C11 and deduced amino acid sequence of the polypeptide (SEQ ID NO:43) enoded by the clone.

[0069]FIG. 23 shows the nucleotide sequence (SEQ ID NO: 44) of the cloneP0184_D11.

[0070]FIG. 24 shows the nucleotide sequence (SEQ ID NO: 45) of the cloneP0240_E05 and deduced amino acid sequence of the polypeptide (SEQ ID NO:46) enoded by the clone.

[0071]FIG. 25 shows the nucleotide sequence (SEQ ID NO: 47) of the cloneP0241_E12.

[0072]FIG. 26 shows the nucleotide sequence (SEQ ID NO: 48) of the cloneP0245_D06.

[0073]FIG. 27 shows the nucleotide sequence (SEQ ID NO: 49) of the cloneP0246_D12.

[0074]FIG. 28 shows the nucleotide sequence (SEQ ID NO: 50) of the cloneP0247_A04 and deduced amino acid sequence of the polypeptide (SEQ ID NO:51) enoded by the clone.

[0075]FIG. 29 shows the nucleotide sequence (SEQ ID NO: 52) of the cloneP0248_B04 and deduced amino acid sequence of the polypeptide (SEQ ID NO:53) enoded by the clone. The underlined amino acid residues at theN-terminal end represent a putative signal peptide.

[0076]FIG. 30 shows the nucleotide sequence (SEQ ID NO: 54 of the cloneP0249_F09 and deduced amino acid sequence of the polypeptide (SEQ ID NO:55) enoded by the clone.

[0077]FIG. 31 shows the nucleotide sequence (SEQ ID NO: 56) of the cloneP0258_A10 and deduced amino acid sequence of the polypeptide (SEQ ID NO:57) enoded by the clone.

[0078]FIG. 32 shows the nucleotide sequence (SEQ ID NO: 58) of the cloneP0262_C10 and deduced amino acid sequence of the polypeptide (SEQ ID NO:59) enoded by the clone.

[0079]FIG. 33 shows the nucleotide sequence (SEQ ID NO: 60) of the cloneP0263 G06.

[0080]FIG. 34 shows the nucleotide sequence (SEQ ID NO: 61) of the cloneP0267_F08.

[0081]FIG. 35 shows the nucleotide sequence (SEQ ID NO: 62) of the cloneP0269_H08 and deduced amino acid sequence of the polypeptide (SEQ ID NO:63) enoded by the clone.

[0082]FIG. 36 shows the nucleotide sequence (SEQ ID NO: 64) of the cloneP0312_C04.

[0083]FIG. 37 shows the nucleotide sequence (SEQ ID NO: 65) of the cloneP0324_H02.

[0084]FIG. 38 shows the nucleotide sequence (SEQ ID NO: 66) of the cloneP0628_H02 and deduced amino acid sequence of the polypeptide (SEQ ID NO:67) enoded by the clone.

[0085]FIG. 39 shows the nucleotide sequence (SEQ ID NO: 68) of the cloneP0629_C08 and deduced amino acid sequence of the polypeptide (SEQ ID NO:69) enoded by the clone.

[0086]FIG. 40 shows the nucleotide sequence (SEQ ID NO: 70) of the cloneP0634_G11.

[0087]FIG. 41 shows the nucleotide sequence (SEQ ID NO: 71) of the cloneP0641_G11 and deduced amino acid sequence of the polypeptide (SEQ ID NO:72) enoded by the clone.

[0088]FIG. 42 shows the nucleotide sequence (SEQ ID NO: 73) of the cloneP0648_E12 and deduced amino acid sequence of the polypeptide (SEQ ID NO:74) enoded by the clone.

[0089]FIG. 43 shows the nucleotide sequence (SEQ ID NO: 75) of the cloneP0697_C03 and deduced amino acid sequence of the polypeptide (SEQ ID NO:76) enoded by the clone.

[0090]FIG. 44 shows the results of differential expression of clonesP00184_D11, P00185_D11, P00188_D12, P00188_E01, P00194_G01, P00194_G05,P00194_H10, P00199_D08, P00203_D04, P00203_E06, P00209_F06, P00219_D02,P00219_F06, P00220_H05, P00222_G03, P00223_F07, P00225_C01, P00227_D11,P00228_F03, P00233_H08, P00235_G08, P00239_C11, P00240_B04, P00240_E05,P00241_E12, P00245_D06, P00246_D12, P00247_A04, P00248_B04, P00249_F09,P00258_A10, P00262_C10, P00263_G06, P00267_F08, P00269_H08, P00312_C04,P00324_H02, P00628_H02, P00629_C08, P00634_G11, P00641_G11, P00648_E12,and P00697_C03 in various heart and kidney disease models in the rat.

DETAILED DESCRIPTION OF THE INVENTION

[0091] A. Definitions

[0092] Unless defined otherwise, technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention belongs. Singleton et al.,Dictionary of Microbiology and Molecular Biology 2^(nd) ed., J. Wiley &Sons (New York, N.Y. 1994), and March, Advanced Organic ChemistryReactions, Mechanisms and Structure 4^(th) ed., John Wiley & Sons (NewYork, N.Y. 1992), provide one skilled in the art with a general guide tomany of the terms used in the present application.

[0093] One skilled in the art will recognize many methods and materialssimilar or equivalent to those described herein, which could be used inthe practice of the present invention. Indeed, the present invention isin no way limited to the methods and materials described. For purposesof the present invention, the following terms are defined below.

[0094] The term “polynucleotide”, when used in singular or plural,generally refers to any polyribonucleotide or polydeoxribonucleotide,which may be unmodified RNA or DNA or modified RNA or DNA. Thus, forinstance, polynucleotides as defined herein include, without limitation,single- and double-stranded DNA, DNA including single- anddouble-stranded regions, single- and double-stranded RNA, and RNAincluding single- and double-stranded regions, hybrid moleculescomprising DNA and RNA that may be single-stranded or, more typically,double-stranded or include single- and double-stranded regions. Inaddition, the term “polynucleotide” as used herein refers totriple-stranded regions comprising RNA or DNA or both RNA and DNA. Thestrands in such regions may be from the same molecule or from differentmolecules. The regions may include all of one or more of the molecules,but more typically involve only a region of some of the molecules. Oneof the molecules of a triple-helical region often is an oligonucleotide.The term “polynucleotide” specifically includes DNAs and RNAs thatcontain one or more modified bases. Thus, DNAs or RNAs with backbonesmodified for stability or for other reasons are “polynucleotides” asthat term is intended herein. Moreover, DNAs or RNAs comprising unusualbases, such as inosine, or modified bases, such as tritylated bases, areincluded within the term “polynucleotides” as defined herein. Ingeneral, the term “polynucleotide” embraces all chemically,enzymatically and/or metabolically modified forms of unmodifiedpolynucleotides, as well as the chemical forms of DNA and RNAcharacteristic of viruses and cells, including simple and complex cells.

[0095] The term “oligonucleotide” refers to a relatively shortpolynucleotide, including, without limitation, single-strandeddeoxyribonucleotides, single- or double-stranded ribonucleotides,RNA:DNA hybrids and double-stranded DNAs. Oligonucleotides, such assingle-stranded DNA probe oligonucleotides, are often synthesized bychemical methods, for example using automated oligonucleotidesynthesizers that are commercially available. However, oligonucleotidescan be made by a variety of other methods, including in vitrorecombinant DNA-mediated techniques and by expression of DNAs in cellsand organisms.

[0096] The term “polypeptide”, in singular or plural, is used herein torefer to any peptide or protein comprising two or more amino acidsjoined to each other in a linear chain by peptide bonds. As used herein,the term refers to both short chains, which also commonly are referredto in the art as peptides, oligopeptides and oligomers, and to longerchains, commonly referred to in the art as proteins. Polypeptides, asdefined herein, may contain amino acids other than the 20 naturallyoccurring amino acids, and may include modified amino acids. Themodification can be anywhere within the polypeptide molecule, such as,for example, at the terminal amino acids, and may be due to naturalprocesses, such as processing and other post-translationalmodifications, or may result from chemical and/or enzymatic modificationtechniques which are well known to the art. The known modificationsinclude, without limitation, acetylation, acylation, ADP-ribosylation,amidation, covalent attachment of flavin, covalent attachment of a hememoiety, covalent attachment of a nucleotide or nucleotide derivative,covalent attachment of a lipid or lipid derivative, covalent attachmentof phosphotidylinositol, cross-linking, cyclization, disulfide bondformation, demethylation, formation of covalent cross-links, formationof cystine, formation of pyroglutamate, formulation,gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation,iodination, methylation, myristoylation, oxidation, proteolyticprocessing, phosphorylation, prenylation, racemization, selenoylation,sulfation, transfer-RNA mediated addition of amino acids to proteinssuch as arginylation, and ubiquitination. Such modifications are wellknown to those of skill and have been described in great detail in thescientific literature, such as, for instance, Creighton, T. E.,Proteins-Structure And Molecular Properties, 2^(nd) Ed., W. H. Freemanand Company, New York (1993); Wold, F., “Posttranslational ProteinModifications: Perspectives and Prospects,” in PosttranslationalCovalent Modification of Proteins, Johnson, B. C., ed., Academic Press,New York (1983), pp. 1-12; Seifter et al., “Analysis for proteinmodifications and nonprotein cofactors,” Meth. Enzymol. 182:626-646(1990), and Rattan et al., Ann. N.Y Acad. Sci. 663:48-62 (1992).

[0097] Modifications can occur anywhere in a polypeptide, including thepeptide backbone, the amino acid side-chains and the amino or carboxyltermini. In fact, blockage of the amino or carboxyl group in apolypeptide, or both, by a covalent modification, is common in naturallyoccurring and synthetic polypeptides and such modifications may bepresent in polypeptides of the present invention, as well. For instance,the amino terminal residue of polypeptides made in E. coli, prior toproteolytic processing, almost invariably will be N-formyhnethionine.

[0098] Modifications that occur in a polypeptide often will be afunction of how the polypeptide is made. For polypeptides made byexpressing a cloned gene in a host, for instance, the nature and extentof the modifications in large part will be determined by the host cellposttranslational modification capacity and the modification signalspresent in the polypeptide amino acid sequence. For instance, it is wellknown that glycosylation usually does not occur in certain bacterialhosts such as E. coli. Accordingly, when glycosylation is desired, apolypeptide is expressed in a glycosylating host, generally eukaryotichost cells. Insect cells often carry out the same posttranslationalglycosylations as mammalian cells and, for this reason, insect cellexpression systems have been developed to express efficiently mammalianproteins having native patterns of glycosylation.

[0099] It will be appreciated that the same type of modification may bepresent in the same or varying degree at several sites in a givenpolypeptide. Also, a given polypeptide may contain many types ofmodifications.

[0100] It will be appreciated that polypeptides are not always entirelylinear. For instance, polypeptides may be branched as a result ofubiquitination, and they may be circular, with or without branching,generally as a result of posttranslational events, including naturalprocessing and events brought about by human manipulation which do notoccur naturally. Circular, branched and branched circular polypeptidesmay be synthesized by non-translation natural process and by entirelysynthetic methods, as well. Such structures are within the scope of thepolypeptides as defined herein.

[0101] The term “amino acid sequence variant” refers to molecules withsome differences in their amino acid sequences as compared to areference (e.g. native sequence) polypeptide. The amino acid alterationsmay be substitutions, insertions, deletions or any desired combinationsof such changes in a native amino acid sequence.

[0102] Substitutional variants are those that have at least one aminoacid residue in a native sequence removed and a different amino acidinserted in its place at the same position. The substitutions may besingle, where only one amino acid in the molecule has been substituted,or they may be multiple, where two or more amino acids have beensubstituted in the same molecule.

[0103] Insertional variants are those with one or more amino acidsinserted immediately adjacent to an amino acid at a particular positionin a native amino acid sequence. Immediately adjacent to an amino acidmeans connected to either the α-carboxy or α-amino functional group ofthe amino acid.

[0104] Deletional variants are those with one or more amino acids in thenative amino acid sequence removed. Ordinarily, deletional variants willhave one or two amino acids deleted in a particular region of themolecule.

[0105] The amino acid sequence variants within the scope of the presentinvention may contain amino acid alterations, including substitutionsand/or insertions and/or deletions in any region of the polypeptide ofSEQ ID NO: 1, including the N- and C-terminal regions. The amino acidsequence variants of the present invention show at least about 75%, morepreferably at least about 85%, even more preferably at least about 90%,most preferably at least about 95% amino acid sequence identity with apolypeptide of SEQ ID NO: 1 or with a native homologue thereof inanother mammalian species, including humans.

[0106] “Sequence identity” is defined as the percentage of amino acidresidues in a candidate sequence that are identical with the amino acidresidues in a native polypeptide sequence, after aligning the sequencesand introducing gaps, if necessary, to achieve the maximum percentsequence identity, and not considering any conservative substitutions aspart of the sequence identity. The % sequence identity values aregenerated by the NCBI BLAST2.0 software as defined by Altschul et al.,(1997), “Gapped BLAST and PSI-BLAST: a new generation of proteindatabase search programs”, Nucleic Acids Res., 25:3389-3402. Theparameters are set to default values, with the exception of the Penaltyfor mismatch, which is set to −1.

[0107] “Stringent” hybridization conditions are sequence dependent andwill be different with different environmental parameters (e.g., saltconcentrations, and presence of organics). Generally, stringentconditions are selected to be about 5° C. to 20° C. lower than thethermal melting point (T_(m)) for the specific nucleic acid sequence ata defined ionic strength and pH. Preferably, stringent conditions areabout 5° C. to 10° C. lower than the thermal melting point for aspecific nucleic acid bound to a complementary nucleic acid. The T_(m)is the temperature (under defined ionic strength and pH) at which 50% ofa nucleic acid (e.g., tag nucleic acid) hybridizes to a perfectlymatched probe

[0108] “Stringent” wash conditions are ordinarily determined empiricallyfor hybridization of each set of tags to a corresponding probe array.The arrays are first hybridized (typically under stringent hybridizationconditions) and then washed with buffers containing successively lowerconcentrations of salts, or higher concentrations of detergents, or atincreasing temperatures until the signal to noise ratio for specific tonon-specific hybridization is high enough to facilitate detection ofspecific hybridization. Stringent temperature conditions will usuallyinclude temperatures in excess of about 30° C., more usually in excessof about 37° C., and occasionally in excess of about 45° C. Stringentsalt conditions will ordinarily be less than about 1000 mM, usually lessthan about 500 mM, more usually less than about 400 mM, typically lessthan about 300 mM, preferably less than about 200 mM, and morepreferably less than about 150 mM. However, the combination ofparameters is more important than the measure of any single parameter.See, e.g., Wetmur et al., J. Mol. Biol. 31:349-70 (1966), and Wetrnur,Critical Reviews in Biochemistry and Molecular Biology 26(34):227-59(1991). In a preferred embodiment, “stringent conditions” or “highstringency conditions,” as defined herein, may be hybridization in 50%formamide, 5× SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodiumphosphate (pH 6.8), 0.1% sodium pyrophosphate, 5× Denhardt's solution,sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10% dextran sulfateat 42° C., with washes at 42° C. in 0.2× SSC (sodium chloride/sodiumcitrate) and 50% formamide at 55° C., followed by a high-stringency washconsisting of 0.1× SSC containing EDTA at 55° C.

[0109] As used herein, the term “polynucleotide encoding a polypeptide”and grammatical equivalents thereof, encompass polynucleotides whichinclude a sequence encoding a polypeptide of the present invention,including polynucleotides that comprise a single continuous region ordiscontinuous regions encoding the polypeptide (for example, interruptedby introns) together with additional regions, that also may containcoding and/or non-coding sequences.

[0110] “Antisense oligodeoxynucleotides” or “antisense oligonucleotides”(which terms are used interchangeably) are defined as nucleic acidmolecules that can inhibit the transcription and/or translation oftarget genes in a sequence-specific manner. The term “antisense” refersto the fact that the nucleic acid is complementary to the coding(“sense”) genetic sequence of the target gene. Antisenseoligonucleotides hybridize in an antiparallel orientation to nascentmRNA through Watson-Crick base-pairing. By binding the target mRNAtemplate, antisense oligonucleotides block the successful translation ofthe encoded protein. The term specifically includes antisense agentscalled “ribozymes” that have been designed to induce catalytic cleavageof a target RNA by addition of a sequence that has natural self-splicingactivity (Warzocha and Wotowiec, “Antisense strategy: biological utilityand prospects in the treatment of hematological malignancies.” Leuk.Lvmnhoma 24:267-281 [1997]).

[0111] The terms “vector”, “polynucleotide vector”, “construct” and“polynucleotide construct” are used interchangeably herein. Apolynucleotide vector of this invention may be in any of several forms,including, but not limited to, RNA, DNA, RNA encapsulated in aretroviral coat, DNA encapsulated in an adenovirus coat, DNA packaged inanother viral or viral-like form (such as herpes simplex, andadeno-associated virus (AAV)), DNA encapsulated in liposomes, DNAcomplexed with polylysine, complexed with synthetic polycationicmolecules, conjugated with transferrin, complexed with compounds such aspolyethylene glycol (PEG) to immunologically “mask” the molecule and/orincrease half-life, or conjugated to a non-viral protein. Preferably,the polynucleotide is DNA. As used herein, “DNA” includes not only basesA, T, C, and G, but also includes any of their analogs or modified formsof these bases, such as methylated nucleotides, intemucleotidemodifications such as uncharged linkages and thioates, use of sugaranalogs, and modified and/or alternative backbone structures, such aspolyaanides.

[0112] The term “antagonist” is used in the broadest sense and includesany molecule that partially or fully blocks, inhibits or neutralizes abiological activity exhibited by a polypeptide of the present invention.In a similar manner, the term “agonist” is used in the broadest senseand includes any molecule that mimics a biological activity exhibited bya polypeptide of the present invention, for example, by specificallychanging the function or expression of such polypeptide, or theefficiency of signaling through such polypeptide, thereby altering(increasing or inhibiting) an already existing biological activity ortriggering a new biological activity.

[0113] The term “recombinant” when used with reference to a cell,animal, or virus indicates that the cell, animal, or virus encodes aforeign DNA or RNA. For example, recombinant cells optionally expressnucleic acids (e.g., RNA) not found within the native (non-recombinant)form of the cell.

[0114] The term “antibody” is used in the broadest sense andspecifically covers monoclonal antibodies (including agonist,antagonist, and neutralizing antibodies), polyclonal antibodies,multi-specific antibodies (e.g., bispecific antibodies), as well asantibody fragments. The monoclonal antibodies specifically include“chimeric” antibodies in which a portion of the heavy and/or light chainis identical with or homologous to corresponding sequences in antibodiesderived from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the chain(s) is identical withor homologous to corresponding sequences in antibodies derived fromanother species or belonging to another antibody class or subclass, aswell as fragments of such antibodies, so long as they exhibit thedesired biological activity (U.S. Pat. No. 4,816,567; Morrison et al.,Proc. Natl. Acad. Sci. USA 81:6851-6855 [1984]). The monoclonalantibodies further include “humanized” antibodies or fragments thereof(such as Fv, Fab, Fab′, F(ab′)₂ or other antigen-binding subsequences ofantibodies) which contain minimal sequence derived from non-humanimmunoglobulin. For the most part, humanized antibodies are humanimmunoglobulins (recipient antibody) in which residues from a CDR of therecipient are replaced by residues from a CDR of a non-human species(donor antibody) such as mouse, rat or rabbit having the desiredspecificity, affinity, and capacity. In some instances, Fv FR residuesof the human immunoglobulin are replaced by corresponding non-humanresidues. Furthermore, humanized antibodies may comprise residues whichare found neither in the recipient antibody nor in the imported CDR orframework sequences. These modifications are made to further refine andmaximize antibody performance. In general, the humanized antibody willcomprise substantially all of at least one, and typically two, variabledomains, in which all or substantially all of the CDR regions correspondto those of a non-human immunoglobulin and all or substantially all ofthe FR regions are those of a human immunoglobulin sequence. Thehumanized antibody optimally also will comprise at least a portion of animmunoglobulin constant region (Fc), typically that of a humanimmunoglobulin. For further details, see Jones et al., Nature,321:522-525 (1986); and Reichmann et al., Nature, 332:323-329 (1988).The humanized antibody includes a PRIMATIZEDO antibody wherein theantigen-binding region of the antibody is derived from an antibodyproduced by immunizing macaque monkeys with the antigen of interest.

[0115] “Antibody fragments” comprise a portion of an intact antibody,preferably the antigen binding or variable region of the intactantibody. Examples of antibody fragments include Fab, Fab′, F(ab′)₂, andFv fragments; diabodies; linear antibodies (Zapata et aL, Protein Eng.8(10):1057-1062 (1995)); single-chain antibody molecules; andmultispecific antibodies formed from antibody fragments.

[0116] The terms “differentially expressed gene,” “differential geneexpression” and their synonyms, which are used interchangeably, refer toa gene whose expression is activated to a higher or lower level in asubject suffering from a disease, specifically a cardiac, kidney orinflammatory disease state, relative to its expression in a normal orcontrol subject. The terms also include genes whose expression isactivated to a higher or lower level at different stages of the samedisease. It is also understood that a differentially expressed gene maybe either activated or inhibited at the nucleic acid level or proteinlevel, or may be subject to alternative splicing to result in adifferent polypeptide product. Such differences may be evidenced by achange in mRNA levels, surface expression, secretion or otherpartitioning of a polypeptide, for example. Differential gene expressionmay include a comparison of expression between two or more genes, or acomparison of the ratios of the expression between two or more genes, oreven a comparison of two differently processed products of the samegene, which differ between normal subjects and subjects suffering from adisease, specifically a cardiac, kidney or inflammatory disease state,or between various stages of the same disease. Differential expressionincludes both quantitative, as well as qualitative, differences in thetemporal or cellular expression pattern in a gene or its expressionproducts among, for example, normal and diseased cells, or among cellswhich have undergone different disease events or disease stages. For thepurpose of this invention, “differential gene expression” is consideredto be present when there is at least an about 1.4-fold, preferably atleast about 1.8-fold, more preferably at least about 2.0-fold, mostpreferably at least about 2.5-fold difference between the expression ofa given gene in normal and diseased subjects, or in various stages ofdisease development in a diseased subject.

[0117] “Cardiac disease” includes congestive heart failure, myocarditis,dilated congestive cardiomyopathy, hypertrophic cardiomyopathy,restrictive cardiomyopathy, mitral valve disease, aortic valve disease,tricuspid valve disease, angina pectoris, myocardial infarction, cardiacarrhythmia, pulmonary hypertension, arterial hypertension, renovascularhypertension, arteriosclerosis, atherosclerosis, and cardiac tumors,along with any disease or disorder that relates to the cardiovascularsystem and related disorders, as well as symptoms indicative of, orrelated to, cardiac disease and related disorders.

[0118] As used herein, “heart failure” refers to an abnormality ofcardiac finction where the heart does not pump blood at the rate neededfor the requirements of metabolizing tissues. The heart failure can becaused by any number of factors, including ischemic, congenital,rheumatic, or idiopathic forms.

[0119] As used herein “congestive heart failure” refers to a syndromecharacterized by left ventricular dysfunction, reduced exercisetolerance, impaired quality of life, and markedly shortened lifeexpectancy. Decreased contractility of the left ventricle leads toreduced cardiac output with consequent systemic arterial and venousvasoconstriction. This vasoconstriction, which appears to be mediated,in part, by the renin-angiotensis system, promotes the vicious cycle offurther reductions of stroke volume followed by an increased elevationof vascular resistance.

[0120] As used herein “infarct” refers to an area of necrosis resultingfrom an insufficiency of blood supply. “Myocardial infarction” refers tomyocardial necrosis resulting from the insufficiency of coronary bloodsupply.

[0121] “Kidney disease” includes acute renal failure,glomerulonephritis, chronic renal failure, azotemia, uremia, immunerenal disease, acute nephritic syndrome, rapidly progressive nephriticsyndrome, nephrotic syndrome, Berger's Disease, chronicnephritic/proteinuric syndrome, tubulointerstital disease, nephrotoxicdisorders, renal infarction, atheroembolic renal disease, renal corticalnecrosis, malignant nephroangiosclerosis, renal vein thrombosis, renaltubular acidosis, renal glucosuria, nepbrogenic diabetes insipidus,Bartter's Syndrome, Liddle's Syndrome, polycystic kidney disease,medullary cystic disease, medullary sponge kidney, hereditary nephritis,and nail-patella syndrome, along with any disease or disorder thatrelates to the renal system and related disorders, as well as symptomsindicative of, or related to, renal or kidney disease and relateddisorders.

[0122] The phrases “polycystic kidney disease” “PKD” and “polycysticrenal disease” are used interchangeably, and refer to a group ofdisorders characterized by a large number of cysts distributedthroughout dramatically enlarged kidneys. The resultant cyst developmentleads to impairment of kidney function and can eventually cause kidneyfailure. “PKD” specifically includes autosomal dominant polycystickidney disease (ADPKD) and recessive autosomal recessive polycystickidney disease (ARPKD), in all stages of development, regardless of theunderlying cause.

[0123] “Inflammatory disease” includes myocarditis, asthma, chronicinflammnation, autoimmune diabetes, tumor angiogenesis, rheumatoidarthritis (RA), rheumatoid spondylitis, osteoarthritis, gouty arthritisand other arthritic conditions, sepsis, septic shock, endotoxic shock,Gram-negative sepsis, toxic shock syndrome, asthma, adult respiratorydistress syndrome, stroke, reperfusion injury, CNS injuries such asneural trauma and ischemia, psoriasis restenosis, cerebral malaria,chronic pulmonary inflammatory disease, silicosis, pulmonary sarcosis,bone resorption diseases such as osteoporosis, graft versus hostreaction, Crohn's Disease, ulcerative colitis including inflammatorybowel disease (IBD), Alzheimer's disease, and pyresis, along with anydisease or disorder that relates to inflammation and related disorders,as well as symptoms indicative of, or related to, inflammation andrelated disorders.

[0124] The terms “treat” or “treatment” refer to both therapeutictreatment and prophylactic or preventative measures, wherein the objectis to prevent or slow down (lessen) an undesired physiological change ordisorder. For purposes of this invention, beneficial or desired clinicalresults include, but are not limited to, alleviation of symptoms,diminishment of extent of disease, stabilized (i.e., not worsening)state of disease, delay or slowing of disease progression, ameliorationor palliation of the disease state, and remission (whether partial ortotal), whether detectable or undetectable. “Treatment” can also meanprolonging survival as compared to expected survival if not receivingtreatment. Those in need of treatment include those already with thecondition or disorder as well as those prone to have the condition ordisorder or those in which the condition or disorder is to be prevented.

[0125] “Chronic” administration refers to administration of the agent(s)in a continuous mode as opposed to an acute mode, so as to maintain thedesired effect for an extended period of time.

[0126] “Intermittent” administration is treatment that is notconsecutively done without interruption, but rather is cyclic in nature.

[0127] Administration “in combination with” one or more furthertherapeutic agents includes simultaneous (concurrent) and consecutiveadministration in any order.

[0128] An “individual” is a vertebrate, preferably a mammal, morepreferably a human.

[0129] “Mammal” for purposes of treatment refers to any animalclassified as a mammal, including humans, domestic and farm animals, andzoo, sports, or pet animals, such as dogs, horses, cats, cows, etc.Preferably, the mammal herein is human.

[0130] An “effective amount” is an amount sufficient to effectbeneficial or desired therapeutic (including preventative) results. Aneffective amount can be administered in one or more administrations.

[0131] “Active” or “activity” means a qualitative biological and/orimmunological property.

[0132] The phrase “immunological property” means immunologicalcross-reactivity with at least one epitope of the reference (nativesequence) polypeptide molecule, wherein, “immunologicalcross-reactivity” means that the candidate polypeptide is capable ofcompetitively inhibiting the qualitative biological activity of thereference (native sequence) polypeptide. The immunologicalcross-reactivity is preferably “specific”, which means that the bindingaffinity of the immunologically cross-reactive molecule identified tothe corresponding polypeptide is significantly higher (preferably atleast about 2 times, more preferably at least about 4-times, mostpreferably at least about 6-times higher) than the binding affinity ofthat molecule to any other known native polypeptide.

[0133] “Carriers” as used herein include pharmaceutically acceptablecarriers, excipients, or stabilizers which are nontoxic to the cell ormammal being exposed thereto at the dosages and concentrations employed.Often the physiologically acceptable carrier is an aqueous pH bufferedsolution. Examples of physiologically acceptable carriers includebuffers such as phosphate, citrate, and other organic acids;antioxidants including ascorbic acid; low molecular weight (less thanabout 10 residues) polypeptide; proteins, such as serum albumin,gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mannose, or dextrins; chelatingagents such as EDTA; sugar alcohols such as mannitol or sorbitol;salt-forming counterions such as sodium; and/or nonionic surfactantssuch as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0134] B. Modes of Carrying Out the Invention

[0135] The practice of the present invention will employ, unlessotherwise indicated, conventional techniques of molecular biology(including recombinant techniques), microbiology, cell biology,biochemistry and immunology, which are within the skill of the art. Suchtechniques are explained fully in the literature, such as, “MolecularCloning: A Laboratory Manuar”, 2^(nd) edition (Sambrook et al., 1989);“Oligonucleotide Synthesis” (M. J. Gait, ed., 1984); “Animal CellCulture” (R. I. Freshney, ed., 1987); “Methods in Enzymology” (AcademicPress, Inc.); “Handbook of Experimental Immunology”, 4^(th) edition (D.M. Weir & C. C. Blackwell, eds., Blackwell Science Inc., 1987); “GeneTransfer Vectors for Mammalian Cells” (J. M. Miller & M. P. Calos, eds.,1987); “Current Protocols in Molecular Biology” (F. M. Ausubel et al.,eds., 1987); “PCR: The Polymerase Chain Reaction”, (Mullis et al., eds.,1994); and “Current Protocols in Immunology” (J. E. Coligan et al.,eds., 1991).

[0136] 1. Identification of Differential Gene Expression and FurtherCharacterization of Differentially Expressed Genes

[0137] The present invention is based on the identification of genesthat are differentially expressed in the left ventricle in theMyocardial Infarction Model, as described in the Examples. Such modelsof differential gene expression can be utilized, among other things, forthe identification of genes which are differentially expressed in normalcells versus cells in a disease state, specifically cardiac, kidney orinflammatory disease state, in cells within different diseases, amongcells within a single given disease state, in cells within differentstages of a disease, or in cells within different time stages of adisease.

[0138] Once a particular differentially expressed gene has beenidentified through the use of one model, its expression pattern can befurther characterized, for example, by studying its expression in adifferent model. A gene may be regulated one way, i.e., the gene canexhibit one differential gene expression pattern, in a given model, butcan be regulated differently in another model. The use, therefore, ofmultiple models can be helpful in distinguishing the roles and relativeimportance of particular genes in a disease, specifically cardiac,kidney or inflammatory disease.

[0139] a. In Vitro Models of Differential Gene Expression

[0140] A suitable model that can be utilized within the context of thepresent invention to discover differentially expressed genes is the invitro specimen model. In a preferred embodiment, the specimen model usesbiological samples from subjects, e.g., peripheral blood, cells andtissues, including surgical and biopsy specimens. Such specimens canrepresent normal peripheral blood and tissue or peripheral blood andtissue from patients suffering from a disease, specifically cardiac,kidney or inflammatory disease, or having undergone surgical treatmentfor disorders involving a disease, such as, for example, coronary bypasssurgery. Surgical specimens can be procured under standard conditionsinvolving freezing and storing in liquid nitrogen (see Karmali et al.,Br. J. Cancer 48:689-96 [1983]). RNA from specimen cells is isolated by,for example, differential centrifugation of homogenized tissue, andanalyzed for differential expression relative to other specimen cells,preferably using microarray analysis.

[0141] Cell lines can also be used to identify genes that aredifferentially expressed in a disease, specifically cardiac, kidney orinflammatory disease. Differentially expressed genes are detected, asdescribed herein, by comparing the pattern of gene expression betweenthe experimental and control conditions. In such models, geneticallymatched disease cell lines (e.g., variants of the same cell line) may beutilized. For example, the gene expression pattern of two variant celllines can be compared, wherein one variant exhibits characteristics ofone disease state while the other variant exhibits characteristics ofanother disease state.

[0142] Alternatively, two variant cell lines, both of which exhibitcharacteristics of the same disease, specifically cardiac, kidney orinflammatory disease, but which exhibit differing degrees of diseasedisorder severity may be used. Further, genetically matched cell linescan be utilized, one of which exhibits characteristics of a disease,specifically cardiac, kidney or inflammatory disease, state, while theother exhibits a normal cellular phenotype. In accordance with thisaspect of the invention, the cell line variants are cultured underappropriate conditions, harvested, and RNA is isolated and analyzed fordifferentially expressed genes, as with the other models. In a preferredembodiment, microarray analysis is used.

[0143] b. In Vivo Models of differential Gene Expression

[0144] In the in vivo model, animal models of a disease, specificallycardiac, kidney or inflammatory disease, and related disorders, can beutilized to discover differentially expressed gene sequences. The invivo nature of such disease models can prove to be especially predictiveof the analogous responses in living patients, particularly humanpatients. Animal models for a disease, specifically cardiac, kidney orinflammatory disease, which can be utilized for in vivo models includeany of the animal models described below. In a preferred embodiment, RNAfrom both the normal and disease state model is isolated and analyzedfor differentially expressed genes using microarray analysis.

[0145] As presented in the examples, three representative in vivocardiac disease models, a representative kidney disease model, and arepresentative inflammatory disease model have been successfullyutilized to identify differentially expressed genes, and are believed tobe useful to further characterize the genes and polypeptides of thepresent invention. These genes are expressed at higher or lower levelsin the disease state, relative to the normal state, and preferably areexpressed at least about a two-fold higher or lower level relative tothe normal state at at least one time point.

[0146] Representative in vivo animal models for use in the presentinvention include the following: generalinflammation—carrageenan-induced paw edema, arachidonic acid-induced earinflammation; arthritis—adjuvant-induced polyarthritis, collagen-inducedarthritis, streptococcal cell wall-induced arthritis; multiplesclerosis—experimental autoimmune encephalomyelitis (EAE); SystemicLupus Erythematosis (SLE); NZB—spontaneous SLE mouse, DNA/anti-DNAimmune complex-induced SLE; insulin-dependent diabetes mellitus—NODspontaneous diabetes mouse; inflammatory bowel disease—acetic acid ortrinitrobenzene sulfonic (TNBS)-induced ulcerative colitis; respiratorydisease—antigen-induced bronchoconstriction (asthma), lipopolysaccharide(LPS)-induced acute respiratory distress syndrome (ARDS);analgesia—acetic acid-induced or phenylquinone-induced writhing, latencyof tail-withdrawal (hot plate); transplant organ rejection—allograftrejection (kidney, lung, heart)-acute and chronic arteriolsclerosis;kidney disease—unilateral nephrectomy (acute renal failure),cyclosporin-induced nephropathy, accelerated crescentic anti-glomerularbasement membrane (GBM) glomerulonephritis, soluble immunecomplex-induced nephritis (see generally Aziz, Bioassavs 17:8 703-12[1995]); and cardiac disease—spontaneous cardiomyopathic hamsters (heartfailure), myocardial infarction (MI) model. pacing-induced model offailure (Riegger model), arrhythmias following myocardial infarction(Harris model), aconitine/chloroform-induced arrhythmisa, carotid arteryinjury (restenosis), balloon angioplasty (restenosis). One skilled inthe art understands that the present invention is not limited to the invivo models recited above and that any known models can be used withinthe context of the present invention.

[0147] C. Microarray Technique

[0148] In a preferred embodiment of the present invention, microarraysare utilized to assess differential expression of genes. In one aspectof the present invention, DNA microarrays are utilized to assess theexpression profile of genes expressed in normal subjects and subjectssuffering from a disease, specifically cardiac, kidney or inflammatorydisease. Identification of the differentially expressed disease genescan be performed by: constructing normalized and subtracted cDNAlibraries from mRNA extracted from the cells or tissue of healthyanimals and an animal model of disease or of healthy patients anddiseased patients, for example, using any of the in vitro or in vivomodels described above; purifying the DNA of cDNA libraries of clonesrepresenting healthy and diseased cells or tissue, microarraying thepurified DNA for expression analysis; and probing microarrays toidentify the genes from the clones that are differentially expressedusing labeled cDNA from healthy and diseased cells or tissues.

[0149] In a specific embodiment of the microarray technique, PCRamplified inserts of cDNA clones are applied to a substrate in a densearray. Preferably at least 10,000 nucleotide sequences are applied tothe substrate. The microarrayed genes, immobilized on the microchip at10,000 elements each, are suitable for hybridization under stringentconditions. Fluorescently labeled cDNA probes may be generated throughincorporation of fluorescent nucleotides by reverse transcription of RNAextracted from tissues of interest. Labeled cDNA probes applied to thechip hybridize with specificity to each spot of DNA on the array. Afterstringent washing to remove non-specifically bound probes, the chip isscarmed by confocal laser microscopy. Quantitation of hybridization ofeach arrayed element allows for assessment of corresponding MRNAabundance. With dual color fluorescence, separately labeled cDNA probesgenerated from two sources of RNA are hybridized pairwise to the array.The relative abundance of the transcripts from the two sourcescorresponding to each specified gene is thus determined simultaneously.The miniaturized scale of the hybridization affords a convenient andrapid evaluation of the expression pattern for large numbers of genes.Such methods have been shown to have the sensitivity required to detectrare transcripts, which are expressed at a few copies per cell, and toreproducibly detect at least approximately two-fold differences in theexpression levels (Schena et al., Proc. Natl. Acad. Sci. USA93(20):106-49 [1996]).

[0150] In a specific embodiment, in vivo models of disease states areused to detect differentially expressed genes. By way of example, threerepresentative cardiac disease models, a representative kidney diseasemodel, and a representative inflammatory disease model were successfullyutilized to identify specific differentially expressed genes.Summarizing the representative general protocol used for such in vivomodels, separate DNA libraries were constructed from niRNA extractedfrom disease state tissue and normal tissue. From these libraries, atleast 20,000 unidentified cDNA clones were preferably chosen foranalysis and microarrayed on chips. Probes generated from normal anddisease tissue, from multiple time points, were hybridized to themicroarray. By this approach, genes, which are differentially expressedin normal and diseased tissue, were revealed and further identified byDNA sequencing. The analysis of the clones for differential expressionreveal genes whose expression is elevated or decreased in associationwith a disease, specifically cardiac, kidney or inflammatory disease, inthe specific in vivo model chosen.

[0151] d. Further Characterization of Differentially Expressed Genes

[0152] The differentially expressed genes of the present invention arescreened to obtain more information about the biological function ofsuch genes. This information can, in turn, lead to the designation ofsuch genes or their gene products as potential therapeutic or diagnosticmolecules, or targets for identifying such molecules.

[0153] The goal of the follow-up work after a differentially expressedgene has been identified is to identify its target cell type(s),function and potential role in disease pathology. To this end, thedifferentially expressed genes are screened to identify cell typesresponding to the gene product, to better understand the mechanism bywhich the identified cell types respond to the gene product, and tofined known signaling pathways that are affected by the expression ofthe gene.

[0154] When further characterization of a differentially expressed geneindicates that a modulation of the gene's expression or a modulation ofthe gene product's activity can inhibit or treat a disease, specificallycardiac, kidney or inflammatory disease, the differentially expressedgene or its gene product becomes a potential drug candidate, or a targetfor developing a drug candidate for the treatment of a cardiac, kidneyor inflammatory disease, or may be used as a diagnostic.

[0155] Where further characterization of a differentially expressed genereveals that modulation of the gene expression or gene product cannotretard or treat a target disease, the differentially expressed gene maystill contribute to developing a gene expression diagnostic patterncorrelative of a disease or its disorders. Accordingly, such genes maybe useful as diagnostics.

[0156] A variety of techniques can be utilized to further characterizethe differentially expressed genes after they are identified.

[0157] First, the nucleotide sequence of the identified genes, which canbe obtained by utilizing standard techniques well known to those ofskill in the art, can be used to further characterize such genes. Forexample, the sequence of the identified genes can reveal homologies toone or more known sequence motifs, which can yield information regardingthe biological function of the identified gene product.

[0158] Second, an analysis of the tissue or cell type distribution ofthe mRNA produced by the identified genes can be conducted, utilizingstandard techniques well known to those of skill in the art. Suchtechniques can include, for example, Northern analyses, microarrays,real time (RT-coupled PCR), and RNase protection techniques. In apreferred embodiment, transcriptional screening is used, which may bebased on the transfection of cells with an inducible promoter-luciferaseplasmid construct, real time PCR, or microarrays, the real time PCR andmicroarray approached being particularly preferred. Such analysesprovide information as to whether the identified genes are expressed infurther tissues expected to contribute to a disease, specificallycardiac, kidney or inflammatory disease. These techniques can alsoprovide quantitative information regarding steady state MRNA regulation,yielding data concerning which of the identified genes exhibits a highlevel of regulation preferably in tissues which can be expected tocontribute to a disease state. Additionally, standard in situhybridization techniques can be utilized to provide informationregarding which cells within a given tissue express the identified gene.Specifically, these techniques can provide information regarding thebiological function of an identified gene relative to a disease,specifically cardiac, kidney or inflammatory disease, where only asubset of the cells within the tissue is thought to be relevant to thedisorder.

[0159] Third, the sequences of the identified differentially expressedgenes can be used, utilizing standard techniques, to place the genesonto genetic maps, e.g., mouse (Copeland et al., Trends in Genetics7:113-18 (1991)) and human genetic maps (Cohen et al., Nature266:698-701 [1993]). This mapping information can yield informationregarding the genes' importance to human disease by identifying genesthat map within genetic regions to which known genetic disease disordersmap.

[0160] After the follow-up screening is completed, relevant, targeted invivo and in vitro systems can be used to more directly assess thebiological function of the identified genes. In vivo systems can includeanimal systems that naturally exhibit symptoms of a disease,specifically cardiac, kidney or inflammatory disease, or ones engineeredto exhibit such symptoms. Animals of any species, including, but notlimited to, mice, rats, rabbits, guinea pigs, pigs, micro-pigs, goats,and non-human primates, e.g. baboons, monkeys, and chimpanzees, can beused to generate animal models of a disease, specifically cardiac,kidney or inflammatory disease. Any technique known in the art can beused to introduce a target gene transgene into animals to produce thefounder lines of transgenic animals. Such techniques include, pronuclearmicroinjection (Hoppe et aL, U.S. Pat. No. 4,873,191 (1989)); retrovirusmediated gene transfer into germ lines (Van der Fatten et al., Proc.Natl. Acad. Sci. USA 82:6148-52 (1985)); gene targeting in embryonicstem cells (Thompson et al., Cell 56:313-21 (1989)); electroporation ofembryos (Lo, Mol. Cell. Biol. 3:1803-14 (1983)); and sperm-mediated genetransfer (Lavitrano et al., Cell 57:717-23 (1989)). For a review of suchtechniques, see Gordon, Intl. Rev. Cytol. 115:171-229 (1989). Furthertechniques will be detailed below, in connection with the gene therapyapplications of the polynucleotides of the present invention.

[0161] The present invention provides for transgenic animals that carrythe transgene in all their cells, as well as animals which carry thetransgene in some, but not all their cells, i.e., mosaic animals. Thetransgene can be integrated, either as a single transgene or inconcatamers, e.g., head-to-head tandems or head-to-tail tandems. Thetransgene can also be selectively introduced into and activated in aparticular cell type by following, for example, the teaching of Lasko etal., Proc. Natl. Acad. Sci. USA 89:6232-36 (1992). The regulatorysequences required for such a cell-type specific activation depends uponthe particular cell type of interest, and will be apparent to those ofskill in the art.

[0162] When it is desired that the transgene be integrated into thechromosomal site of the endogenous target gene, gene targeting ispreferred. Briefly, when such a technique is to be utilized, vectorscontaining some nucleotide sequences homologous to the endogenous targetgene of interest are designed for the purpose of integrating, viahomologous recombination with chromosomal sequences, into and disruptingthe function of the nucleotide sequence of the endogenous target gene.The transgene can also be selectively introduced into a particular celltype, thus inactivating the endogenous gene of interest in only thatcell type, by following the teaching of Gu et al. (Science 265:103-06[1994]). The regulatory sequences required for such a cell-type specificinactivation depends upon the particular cell type of interest, and willbe apparent to those of skill in the art.

[0163] Once transgenic animals have been generated, the expression ofthe recombinant target gene and protein can be assayed using standardtechniques. Initial screening can be accomplished by Southern blotanalysis or PCR techniques to analyze animal tissues to assay whetherintegration of the transgene has taken place. The level of MnRNAexpression of the transgene in the tissues of the transgenic animals canalso be assessed using techniques which include Northern blot analysisof tissue samples obtained from the animal, in situ hybridizationanalysis, and RT-coupled PCR. Samples of target gene-expressing tissuecan also be evaluated immunocytochemically using antibodies specific forthe transgenic product of interest.

[0164] The transgenic animals that express target gene niRNA or targetgene transgene peptide (detected immunocytochemically, using antibodiesdirected against target gene product epitopes) at easily detectablelevels should then be further evaluated to identify those animals whichdisplay disease characteristics or symptoms. Additionally, specific celltypes within the transgenic animals can be analyzed for cellularphenotypes characteristic of a disease, specifically cardiac, kidney orinflarumatory disease. Such cellular phenotypes can include, forexample, differential gene expression characteristic of cells within agiven disease state of interest. Further, such cellular phenotypes caninclude an assessment of a particular cell type diagnostic pattern ofexpression and its comparison to known diagnostic expression profiles ofthe particular cell type in animals exhibiting a disease, specificallycardiac, kidney or inflammatory disease. Such transgenic animals serveas suitable models. Once transgenic founder animals are produced, theycan be bred, inbred, outbred, or crossbred to produce colonies of theparticular animal.

[0165] The animal models described above and in the Examples, can beused to generate cell lines for use in cell-based in vitro assays tofurther characterize the differentially expressed genes of the inventionand their gene products. Techniques that can be used to derive acontinuous cell line from transgenic animals are disclosed, for example,by Small et al., Mol. Cell Biol. 5:642-48 (1985).

[0166] Alternatively, cells of a cell type known to be involved in acardiac, kidney or inflammratory disease can be transfected withsequences capable of increasing or decreasing the amount of target geneexpression within the cell. For example, sequences of the differentiallyexpressed genes herein can be introduced into, and overexpressed in, thegenome of the cell of interest, or if endogenous target gene sequencesare present, they can either be overexpressed or, be disrupted in orderto underexpress or inactivate target gene expression.

[0167] The information obtained through such characterizations cansuggest relevant methods for the treatment of a disease, specificallycardiac, kidney or inflammatory disease, involving the gene of interest.For example, treatment can include a modulation of gene expression orgene product activity. Characterization procedures such as thosedescribed herein can indicate where such modulation should involve anincrease or a decrease in the expression or activity of the gene or geneproduct of interest.

[0168] 2. Production of Polvnucleotides and Polypeptides

[0169] The polypeptides of the present invention are preferably producedby techniques of recombinant DNA technology. DNA encoding a nativepolypeptide herein can be obtained from cDNA libraries prepared fromtissue believed to possess the corresponding mRNA and to express it at adetectable level. For example, cDNA library can be constructed byobtaining polyadenylated mRNA from a cell line known to express thedesired polypeptide, and using the MRNA as a template to synthesizedouble-stranded cDNA. In the present case, a suitable source for thedesired niRNA may be heart tissue obtained from normal heart or from theMyocardial Infarction Model (MI model) mentioned above, and described indetail in the Examples. The polypeptide genes of the present inventioncan also be obtained from a genomic library, such as a human genomiccosmid library.

[0170] Libraries, either cDNA or genomic, are screened with probesdesigned to identify the gene of interest or the protein encoded by it.For cDNA expression libraries, suitable probes include monoclonal andpolyclonal antibodies that recognize and specifically bind to apolypeptide of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24,26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69,72, 74, and 76. For cDNA libraries, suitable probes includeoligonucleotide probes (generally about 20-80 bases) that encode knownor suspected portions of a polypeptide herein, from the same ordifferent species, and/or complementary or homologous cDNAs or fragmentsthereof that encode the same or a similar gene. Appropriate probes forscreening genomic libraries include, without limitation,oligonucleotides, cDNAs, or fragments thereof that encode the same or asimilar gene, and/or homologous genomic DNAs or fragments thereof.Screening the cDNA and genornic libraries with the selected probe may beconducted using standard protocols as described, for example, inChapters 10-12 of Sambrook et al., Molecular Cloning: A LaboratoryManuaL New York, Cold Spring Harbor Laboratory Press (1989).

[0171] According to a preferred method, carefully selectedoligonucleotide probes are used to screen cDNA libraries from varioustissues, preferably from heart and/or kidney tissues. Theoligonucleotide sequences selected as probes should be sufficient inlength and sufficiently unique and unambiguous that false positives areminimized. The actual sequences can be designed based on regions of SEQID NO: 2 which have the least codon redundance. The oligonucleotides maybe degenerate at one or more positions. The use of degenerateoligonucleotides is of particular importance where a library is screenedfrom a species in which preferential codon usage is not known.

[0172] The oligonuleotides must be labeled such that they can bedetected upon hybridization to DNA in the library screened. Preferably,the 5′ end of the oligonucleotide is radiolabeled, using APT (e.g. γ³²P)and polynucleotide kinase. However, other labeling, e.g. biotinylationor enzymatic labeling are also suitable.

[0173] Alternatively, to obtain DNA encoding a homologue of ratpolypeptides specifically disclosed herein in another mammalian species,e.g. in humans, one only needs to conduct hybridization screening withlabeled rat DNA or fragments thereof, selected following the principlesoutlined above, in order to detect clones which contain homologoussequences in the cDNA libraries obtained from appropriate tissues (e.g.heart or kidney) of the particular animal, such as human (cross-specieshybridization). Full-length clones can then be identified, for example,by restriction endonuclease analysis and nucleic acid sequencing. Iffull-length clones are not identified, appropriate fragments arerecovered from the various clones and ligated at restriction sitesconmmon to the fragments to assemble a full-length clone. cDNAs encodingthe polypeptides of the present invention can also be identified andisolated by other known techniques, such as by direct expression cloningor by using the PCR technique, both of which are well known aredescribed in textbooks, such as those referenced hereinbefore.

[0174] Once the sequence is known, the nucleic acid encoding aparticular polypeptide of the present invention can also be obtained bychemical synthesis, following known methods, such as the phosphoramiditemethod (Beaucage and Caruthers, Tetrahedron Letters 22:1859 [1981];Matteucci and Caruthers, Tetrahedron Letters 21:719 [1980]; andMatteucci and Caruthers, J. Amer. Chem. Soc. 103: 3185 [1981]), and thephosphotriester approach (Ito et al., Nucleic Acids Res. 10:1755-1769[1982]).

[0175] The cDNA encoding the desired polypeptide of the presentinvention is inserted into a replicable vector for cloning andexpression. Suitable vectors are prepared using standard techniques ofrecombinant DNA technology, and are, for example, described in thetextbooks cited above. Isolated plasmids and DNA fragments are cleaved,tailored, and ligated together in a specific order to generate thedesired vectors. After ligation, the vector containing the gene to beexpressed is transformed into a suitable host cell.

[0176] Host cells can be any eukaryotic or prokaryotic hosts known forexpression of heterologous proteins.

[0177] The polypeptides of the present invention can be expressed ineukaryotic hosts, such as eukaryotic microbes (yeast), cells isolatedfrom multicellular organisms (martmalian cell cultures), plants andinsect cells.

[0178] While prokaryotic host provide a convenient means to synthesizeeukaryotic proteins, when made this fashion, proteins usually lack manyof the immunogenic properties, three-dimensional conformation,glycosylation, and other features exhibited by authentic eukaryoticproteins. Eukaryotic expression systems overcome these limitations.

[0179] Yeasts are particularly attractive as expression hosts for anumber of reasons. They can be rapidly growth on inexpensive (minimal)media, the recombinant can be easily selected by complementation,expressed proteins can be specifically engineered for cytoplasmiclocalization or for extracellular export, and are well suited forlarge-scale fermentation.

[0180]Saccharomyces cerevisiae is the most commonly used among lowereukaryotic hosts. However, a number of other genera, species, andstrains are also available and useful herein, such as Pichia pastoris(EP 183,070; Sreekrishna et al, J. Basic Microbiol. 28:165-278 [1988]).Yeast expression systems are commercially available, and can bepurchased, for example, from Invitrogen (San Diego, Calif.). Otheryeasts suitable for VEGF expression include, without limitation,Kluyveromyces hosts (U.S. Pat. No. 4,943,529), e.g. Kluyveromyceslactis; Schizosaccharomyces pombe (Beach and Nurse, Nature 290:140(1981); Aspergillus hosts, e.g. A. niger (Kelly and Hynes, EMBO J.4:475-479 [1985]) and A. nidulans (Ballance et al., Biochem. Biophys.Res. Commun. 112:284-289 [1983]), and Hansenula hosts, e.g. Hansenulapolymorpha.

[0181] Preferably a methylotrophic yeast is used as a host in performingthe methods of the present invention. Suitable methylotrophic yeastsinclude, but are not limited to, yeast capable of growth on methanolselected from the group consisting of the genera Pichia and Hansenula. Alist of specific species which are exemplary of this class of yeasts maybe found, for example, in C. Anthony, The Biochemistry of Methvlotrophs,269 (1982). Presently preferred are methylotrophic yeasts of the genusPichia such as the auxotrophic Pichia pastoris GS115 (NRRL Y-15851);Pichia pastoris GS190 (NRRL Y-18014) disclosed in U.S. Pat. No.4,818,700; and Pichia pastoris PPF1 (NRRL Y-18017) disclosed in U.S.Pat. No. 4,812,405. Auxotrophic Pichia pastoris strains are alsoadvantageous to the practice of this invention for their ease ofselection. It is recognized that wild type Pichia pastoris strains (suchas NRRL Y-11430 and NRRL Y-1 1431) may be employed with equal success ifa suitable transforming marker gene is selected, such as the use of SUC2to transform Pichia pastoris to a strain capable of growth on sucrose,or if an antibiotic resistance marker is employed, such as resistance toG418. Pichia pastoris linear plasmids are disclosed, for example, inU.S. Pat. No. 5,665,600.

[0182] Suitable promoters used in yeast vectors include the promotersfor 3-phosphoglycerate kinase (Hitzeman et al., J. Biol. Chem. 255:2073[1980]); and other glycolytic enzymes (Hess et aL, J. Adv. Enzyme Res.7:149 [1968]; Holland et al., Biochemistry 17:4900 [1978]), e.g.,enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyvuratedecarboxylase, phosphoffiuctokinase, glucose-6-phosphate isomerase,3-phosphoglycerate mutase, pyruvate kinase, triosephosphate somerase,phosphoglucose isomerase, and glucokinase. In the constructions ofsuitable expression plasmids, the termination sequences associated withthese genes are also ligated into the expression vector 3′ of thesequence desired to be expressed to provide polyadenylation of the mRNAand termination. Other promoters that have the additional advantage oftranscription controlled by growth conditions are the promoter regionsfor alcohol oxidase 1 (AOX1, particularly preferred for expression inPichia), alcohol dehydrogenase 2, isocytochrome C, acid phosphatase,degradative enzymes associated with nitrogen metabolism, and theaforementioned glyceraldehyde-3-phosphate dehydrogenase, and enzymesresponsible for maltose and galactose utilization. Any plasmid vectorcontaining a yeast-compatible promoter and termination sequences, withor without an origin of replication, is suitable. Yeast expressionsystems are commercially available, for example, from ClontechLaboratories, Inc. (Palo Alto, Cali., e.g. pYEX 4T family of vectors forS. cerevisiae), Invitrogen (Carlsbad, Calif., e.g. pPICZ series EasySelect Pichia Expression Kit) and Stratagene (La Jolla, Calif., e.g.ESP™ Yeast Protein Expression and Purification System for S. pombe andpESC vectors for S. cerevisiae).

[0183] Cell cultures derived from multicellular organisms may also beused as hosts to practice the present invention. While both invertebrateand vertebrate cell cultures are acceptable, vertebrate cell cultures,particularly mammalian cells, are preferable. Examples of suitable celllines include monkey kidney CV1 cell line transformed by SV40 (COS-7,ATCC CRL 1651); human embryonic kidney cell line 293S (Graham et al, J.Gen. Virol. 36:59 [1977]); baby hamster kidney cells (BHK, ATCC CCL 10);Chinese hamster ovary (CHO) cells (Urlaub and Chasin, Proc. Natl. Acad.Sci. USA 77:4216 [1980]; monkey kidney cells (CVI-76, ATCC CCL 70);African green monkey cells (VERO-76, ATCC CRL-1587); human cervicalcarcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL34); human lung cells (W138, ATCC CCL 75); and human liver cells (HepG2, HB 8065).

[0184] Suitable promoters used in mammalian expression vectors are oftenof viral origin. These viral promoters are commonly derived fromcytomeagolavirus (CMV), polyoma virus, Adenovirus2, and Simian Virus 40(SV40). The SV40 virus contains two promoters that are termed the earlyand late promoters. They are both easily obtained from the virus as oneDNA fragment that also contains the viral origin of replication (Fierset al., Nature 273:113 [1978]). Smaller or larger SV40 DNA fragments mayalso be used, provided they contain the approximately 250-bp sequenceextending from the HindIII site toward the BglI site located in theviral origin of replication. An origin of replication may be obtainedfrom an exogenous source, such as SV40 or other virus, and inserted intothe cloning vector. Alternatively, the host cell chromosomal mechanismmay provide the origin of replication. If the vector containing theforeign gene is integrated into the host cell chromosome, the latter isoften sufficient.

[0185] Eukaryotic expression systems employing insect cell hosts mayrely on either plasmid or baculoviral expression systems. The typicalinsect host cells are derived from the fall army worm (Spodopterafrugiperda). For expression of a foreign protein these cells areinfected with a recombinant form of the baculovirus Autographacalifornica nuclear polyhedrosis virus which has the gene of interestexpressed under the control of the viral polyhedrin promoter. Otherinsects infected by this virus include a cell line known commercially as“High 5” (Invitrogen) which is derived from the cabbage looper(Trichoplusia ni). Another baculovirus sometimes used is the Bombyx morinuclear polyhedorsis virus which infect the silk worm (Bombyx mori).Numerous baculovirus expression systems are commercially available, forexample, from Invitrogen (Bac-N-Blue™), Clontech (BacPAK™ BaculovirusExpression System), Life Technologies (BAC-TO-BAC™), Novagen (Bac VectorSystem™), Pharmingen and Quantum Biotechnologies). Another insect cellhost is common fruit fly, Drosophila melanogaster, for which a transientor stable plasmid based transfection kit is offered commercially byInvitrogen (The DES™ System).

[0186] Prokaryotes are the preferred hosts for the initial cloningsteps, and are particularly useful for rapid production of large amountsof DNA, for production of single-stranded DNA templates used forsite-directed mutagenesis, for screening many mutants simultaneously,and for DNA sequencing of the mutants generated. E. coli strainssuitable for the production of the polypeptides of the present inventioninclude, for example, BL21 carrying an inducible T7 RNA polymerase gene(Studier et al., Methods Enzymol. 185:60-98 [1990]); AD494 (DE3); EB105; and CB (E. coli B) and their derivatives; K12 strain 214 (ATCC31,446); W3 110 (ATCC 27,325); X1776 (ATCC 31,537); HB101 (ATCC 33,694);JM101 (ATCC 33,876); NM522 (ATCC 47,000); NM538 (ATCC 35,638); NM539(ATCC 35,639), etc. Many other species and genera of prokaryotes may beused as well. Prokaryotes, e.g. E. coli, produce the polypeptides of thepresent invention in an unglycosylated form.

[0187] Vectors used for transformation of prokaryotic host cells usuallyhave a replication site, marker gene providing for phenotypic selectionin transformed cells, one or more promoters compatible with the hostcells, and a polylinker region containing several restriction sites forinsertion of foreign DNA. Plasmids typically used for transformation ofE. coli include pBR322, pUC18, pUC19, pUC118, pUC119, and BluescriptM13, all of which are commercially available and described in Sections1.12-1.20 of Sambrook et al., supra. The promoters commonly used invectors for the transformation of prokaryotes are the T7 promoter(Studier et al., supra); the tryptophan (trp) promoter (Goeddel et al.,Nature 281:544 [1979]); the alkaline phosphatase promoter (phoA); andthe P-lactamase and lactose (lac) promoter systems. In E. coli, somepolypeptides accumulate in the form of inclusion bodies, and need to besolubilized, purified, and refolded. These steps can be carried out bymethods well known in the art.

[0188] Many eukaryotic proteins, including the polypeptide of SEQ IDNOS: 26 and 53 disclosed herein, contain an endogenous signal sequenceas part of the primary translation product. This sequence targets theprotein for export from the cell via the endoplasmic reticulum and Golgiapparatus. The signal sequence is typically located at the aminoterminus of the protein, and ranges in length from about 13 to about 36amino acids. Although the actual sequence varies among proteins, allknown eukaryotic signal sequences contain at least one positivelycharged residue and a highly hydrophobic stretch of 10-15 amino acids(usually rich in the amino acids leucine, isoleucine, valine andphenylalanine) near the center of the signal sequence. The signalsequence is normally absent from the secreted form of the protein, as itis cleaved by a signal peptidase located on the endoplasmic reticulumduring translocation of the protein into the endoplasmic reticulum. Theprotein with its signal sequence still attached is often referred to asthe pre-protein, or the immature form of the protein, in contrast to theprotein from which the signal sequence has been cleaved off, which isusually referred to as the mature protein. Proteins may also be targetedfor secretion by linking a heterologous signal sequence to the protein.This is readily accomplished by ligating DNA encoding a signal sequenceto the 5′ end of the DNA encoding the protein, and expressing the fusionprotein in an appropriate host cell. Prokaryotic and eukaryotic (yeastand mammalian) signal sequences may be used, depending on the type ofthe host cell. The DNA encoding the signal sequence is usually excisedfrom a gene encoding a protein with a signal sequence, and then ligatedto the DNA encoding the protein to be secreted. Alternatively, thesignal sequence can be chemically synthesized. The signal must befunctional, i.e. recognized by the host cell signal peptidase such thatthe signal sequence is cleaved and the protein is secreted. A largevariety of eukaryotic and prokaryotic signal sequences is known in theart, and can be used in performing the process of the present invention.Yeast signal sequences include, for example, acid phosphatase, alphafactor, alkaline phosphatase and invertase signal sequences. Prokaryoticsignal sequences include, for example LamB, OmpA, OmpB and OmpF, MalE,PhoA, and P lactamase.

[0189] Mammalian cells are usually transformed with the appropriateexpression vector using a version of the calcium phosphate method(Graham et aL, Virology 52:546 [1978]; Sambrook et al., supra, sections16.32-16.37), or, more recently, lipofection . However, other methods,e.g. protoplast fusion, electroporation, direct microinjection, etc. arealso suitable.

[0190] Yeast hosts are generally transformed by the polyethylene glycolmethod (Hinnen, Proc. Natl. Acad, Sci. USA 75:1929 [1978]). Yeast, e.g.Pichia pastoris, can also be transformed by other methodologies, e.g.electroporation.

[0191] Prokaryotic host cells can, for example, be transformed using thecalcium chloride method (Sambrook et al., supra, section 1.82), orelectroporation.

[0192] More recently, techniques have been developed for the expressionof heterologous proteins in the milk of non-human transgenic animals.For example, Krimpenfort et al., Biotechnology 9:844-847 (1991)describes microinjection of fertilized bovine oocytes with genesencoding human proteins and development of the resulting embryos insurrogate mothers. The human genes were fused to thebovine.alpha.S.sub.1 casein regulatory elements. This general technologyis also described in PCT Application W091/08216 published Jun. 13, 1991.PCT application W088/00239, published Jan. 14, 1988, describesprocedures for obtaining suitable regulatory DNA sequences for theproducts of the mammary glands of sheep, including beta lactoglobulin,and the construction of transgenic sheep modified so as to secreteforeign proteins in milk. PCT publication W088/01648, published Mar. 10,1988, generally describes construction of transgenic animals whichsecrete foreign proteins into milk under control of the regulatorysequences of bovine alpha lactalbumin gene. PCT application W088/10118,published Dec. 29, 1988, describes construction of transgenic mice andlarger mammals for the production of various recombinant human proteinsin milk. Thus, techniques for construction of appropriate host vectorscontaining regulatory sequences effective to produce foreign proteins inmammary glands and cause the secretion of said protein into milk areknown in the art.

[0193] Among the milk-specific protein promoters are the caseinpromoters and the beta lactoglobulin promoter. The casein promoters may,for example, be selected from an alpha casein promoter, a beta caseinpromoter or a kappa casein promoter. Preferably, the casein promoter isof bovine origin and is an alpha S-1 casein promoter. Among thepromoters that are specifically activated in mammary is the longterminal repeat (LTR) promoter of the mouse mammary tumor virus (MMTV).The milk-specific protein promoter or the promoters that arespecifically activated in mammary tissue may be derived from either cDNAor genomic sequences. Preferably, they are genomic in origin.

[0194] Signal peptides that are useful in expressing heterologousproteins in the milk of transgenic amammals include milk-specific signalpeptides or other signal peptides useful in the secretion and maturationof eukaryotic and prokaryotic proteins. Preferably, the signal peptideis selected from milk-specific signal peptides or the signal peptide ofthe desired recombinant protein product, if any. Most preferably, themilk-specific signal peptide is related to the milk-specific promoterused in the expression system of this invention.

[0195] The present invention includes amino acid sequence variants ofthe native rat polypeptides specifically disclosed herein or theiranalogues in any other animal, e.g. mammalian species, including humans.Such amino acid sequence variants can be produced by expressing theunderlying DNA sequence in a suitable recombinant host cell, asdescribed above, or by in vitro synthesis of the desired polypeptide.The nucleic acid sequence encoding a polypeptide variant of the presentinvention is preferably prepared by site-directed mutagenesis of thenucleic acid sequence encoding the corresponding native (e.g. human)polypeptide. Particularly preferred is site-directed mutagenesis usingpolymerase chain reaction (PCR) amplification (see, for example, U.S.Pat. No. 4,683,195 issued Jul. 28, 1987; and Current Protocols InMolecular Biology, Chapter 15 (Ausubel et al., ed., 1991). Othersite-directed mutagenesis techniques are also well known in the art andare described, for example, in the following publications: CurrentProtocols In Molecular Biology, supra, Chapter 8; Molecular Cloning: ALaboratory Manual., 2^(nd) edition (Sambrook et al., 1989); Zoller etal., Methods Enzvmol. 100:468-500 (1983); Zoller & Smith, DNA 3:479-488(1984); Zoller et al., Nucl. Acids Res., 10:6487 (1987); Brake et al.,Proc. Natl. Acad. Sci. USA 81:4642-4646 (1984); Botstein et al., Science229:1193 (1985); Kunkel et al., Methods Enzymol. 154:367-82 (1987),Adelman et aL, DNA 2:183 (1983); and Carter et al., Nucl. Acids Res.,13:4331 (1986). Cassette mutagenesis (Wells et al., Gene, 34:315[1985]), and restriction selection mutagenesis (Wells et al., Philos.Trans. R. Soc. London SerA, 317:415 [1986]) may also be used.

[0196] Amino acid sequence variants with more than one amino acidsubstitution may be generated in one of several ways. If the amino acidsare located close together in the polypeptide chain, they may be mutatedsimultaneously, using one oligonucleotide that codes for all of thedesired amino acid substitutions. If, however, the amino acids arelocated some distance from one another (e.g. separated by more than tenamino acids), it is more difficult to generate a single oligonucleotidethat encodes all of the desired changes. Instead, one of two alternativemethods may be employed. In the first method, a separate oligonucleotideis generated for each amino acid to be substituted. The oligonucleotidesare then annealed to the single-stranded template DNA simultaneously,and the second strand of DNA that is synthesized from the template willencode all of the desired amino acid substitutions. The alternativemethod involves two or more rounds of mutagenesis to produce the desiredmutant.

[0197] The amino acid sequence variants of the present invention includepolypeptides in which the membrane spanning (transmembrane) region orregions are deleted or inactivated. Deletion or inactivation of theseportions of the molecule yields soluble proteins, which are no longercapable of membrane anchorage. Inactivation may, for example, beachieved by deleting sufficient residues (but less than the entiretransmembrane region) to produce a substantially hydrophilic hydropathyprofile at this site, or by substituting with heterologous residueswhich accomplish the same result. For example, the transmembraneregion(s) may be substituted by a random or predetermined sequence ofabout 5 to 50 serine, threonine, lysine, arginine, glutamine, asparticacid and like hydrophilic residues, which altogether exhibit ahydrophilic hydropathy profile. Like the transmembrane region deletionalvariants, these variants are “soluble”, i.e. secreted into the culturemedium of recombinant hosts. Soluble variants of the native polypeptidesof the present invention may be used to make fusions at their N- orC-terninus to immunogenic polypeptides, e.g. bacterial polypeptides suchas beta-lactamase or an enzyme encoded by the E. coli trp locus, oryeast protein, and C-termninal fusions with proteins having a longhalf-life such as immunoglobulin regions (preferably immunoglobulinconstant regions to yield imnnmunoadhesins), albumin, or ferritin, asdescribed in WO 89/02922 published on Apr. 6, 1989. For the productionof immunoglobulin fusions see also U.S. Pat. No. 5,428,130 issued Jun.27, 1995.

[0198] 3. Production of Antibodies

[0199] The present invention includes antibodies that specifically binda polypeptide of SEQ ID NO: 2 or another mammalian (e.g. human)homologue of such polypeptide. Such antibodies find utility as reagentsused, for example, in analytical chemistry or process sciences, asdiagnostic and/or therapeutics.

[0200] Methods of preparing polyclonal antibodies are known in the art.Polyclonal antibodies can be raised in a mammal, for example, by one ormore injections of an immunizing agent and, if desired, an adjuvant.Typically, the immunizing agent and/or adjuvant will be injected in themammal by multiple subcutaneous or intraperitoneal injections. It may beuseful to conjugate the immunizing agent to a protein known to beimmunogenic in the mammal being immunized, such as serum albumin, orsoybean trypsin inhibitor. Examples of adjuvants which may be employedinclude Freund's complete adjuvant and MPL-TDM.

[0201] According to one approach, monoclonal antibodies may be preparedusing hybridoma methods, such as those described by Kohler and Milstein,Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, orother appropriate host animal, is typically immunized with an immunizingagent to elicit lymphocytes that produce or are capable of producingantibodies that will specifically bind to the immunizing agent.Alternatively, the lymphocytes may be immunized in vitro. Generally,either peripheral blood lymphocytes (“PBLs”) are used if cells of humanorigin are desired, or spleen cells or lymph node cells are used ifnon-human mammalian sources are desired. The lymphocytes are then fusedwith an immortalized cell line using a suitable fusing agent, such aspolyethylene glycol, to form a hybridoma cell [Goding, MonoclonalAntibodies: Principles and Practice, Academic Press, (1986) pp. 59-103].Immortalized cell lines are usually transformed mammalian cells,particularly myeloma cells of rodent, bovine and human origin. Usually,rat or mouse myeloma cell lines are employed. The hybridoma cells may becultured in a suitable culture medium that preferably contains one ormore substances that inhibit the growth or survival of the unfused,immortalized cells. Preferred immortalized cell lines are those thatfuse efficiently, support stable high level expression of antibody bythe selected antibody-producing cells, and are sensitive to a mediumsuch as HAT medium.

[0202] The culture medium in which the hybridoma cells are cultured canthen be assayed for the presence of monoclonal antibodies directedagainst the particular polypeptide used, such as a rat polypeptide ofSEQ ID NO: 2 or its human homologue. Preferably, the binding specificityof monoclonal antibodies produced by the hybridoma cells is determinedby immunoprecipitation or by an in vitro binding assay, such asradioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchardanalysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0203] After the desired hybridoma cells are identified, the clones maybe subcloned by limiting dilution procedures and grown by standardmethods [Goding, supra]. Suitable culture media for this purposeinclude, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640medium. Alternatively, the hybridoma cells may be grown in vivo asascites in a mammal.

[0204] The monoclonal antibodies secreted by the subclones may beisolated or purified from the culture medium or ascites fluid byconventional immunoglobulin purification procedures such as, forexample, protein A-Sepharose, hydroxylapatite chromatography, gelelectrophoresis, dialysis, or affmity chromatography.

[0205] Alternatively, monoclonal antibodies may be made by recombinantDNA methods, such as those described in U.S. Pat. No. 4,816,567. DNAencoding the monoclonal antibodies of the invention can be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of murine antibodies). The hybridomacells discussed above serve as a preferred source of such DNA. Onceisolated, the DNA may be placed into expression vectors, which are thentransfected into host cells such as COS cells, Chinese hamster ovary(CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells.

[0206] The antibodies, including antibody fragments, such as Fv, Fab,Fab′, F(ab′)₂ or other antigen-binding subsequences of antibodies, maybe humanized. Humanized antibodies contain minimal sequence derived froma non-human immunoglobulin. More specifically, in humanized antibodiesresidues from a complementary determining region (CDR) of a humanimununoglobulin (the recipient) are replaced by residues from a CDR of anon-human species (donor antibody) such as mouse, rat or rabbit havingthe desired specificity, affinity and capacity. In some instances, Fvframework residues of the human immunoglobulin are also replaced bycorresponding non-human residues. Humanized antibodies may additionallycomprise residues that are found neither in the recipient antibody norin the imported CDR or framework sequences [Jones et al., Nature,321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988)].

[0207] Methods for humanizing non-human antibodies are well known in theart. Generally, a humanized antibody has one or more amino acid residuesintroduced into it from a non-human source. These non-human amino acidresidues are often referred to as “import” residues, which are typicallytaken from an “import” variable domain. Humanization can be essentiallyperformed following the method of Winter and co-workers [Jones et al.,Nature 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988);Verhoeyen et al., Science, 239:1534-1536 (1988)], by substituting rodentCDRs or CDR sequences for the corresponding sequences of a humanantibody. In addition, human antibodies can be produced using varioustechniques known in the art, including phage display libraries[Hoogenboom and Winter, J. Mol. Biol, 227:381 (1991); Marks et al., J.Mol. Biol., 222:581 (1991)]. The techniques of Cole et al. and Boerneret al. are also available for the preparation of human monoclonalantibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, AlanR. Liss, p. 77 (1985) and Boemer et al., J. Immunol., 147(1):86-95(1991)]. Similarly, human antibodies can be made by introducing of humanimmunoglobulin loci into transgenic animals, e.g., mice in which theendogenous immunoglobulin genes have been partially or completelyinactivated. Upon challenge, human antibody production is observed,which closely resembles that seen in humans in all respects, includinggene rearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the followingscientific publications: Marks et al., Bio/Technology 10, 779-783(1992); Lonberg et al., Nature 368 856-859 (1994); Morrison, Nature 368,812-13 (1994); Fishwild et aL, Nature Biotechnology 14, 845-51 (1996);Neuberger, Nature Biotechnology 14, 826 (1996); Lonberg and Huszar,Intern. Rev. Immunol. 13 65-93 (1995).

[0208] The antibodies may be bispecific, in which one specificity is forpolypeptide of the present invention, and the other specificity foranother protein, such as, a second polypeptide of the present inventionor another polypeptide.

[0209] 4. Uses

[0210] a. Polynucleotides

[0211] The differentially expressed genes identified in accordance withthe present invention may be used to design specific oligonucleotideprobes and primers. In certain preferred embodiments, the term “primer”as used here includes any nucleic acid capable of primingtemplate-dependent synthesis of a nascent nucleic acid. In certain otherembodiments, the nucleic acid may be able to hybridize a template, butnot be extended for synthesis of nascent nucleic acid that iscomplementary to the template.

[0212] In certain embodiments of the present invention the term“template” may refer to a nucleic acid that is used in the creation of acomplementary nucleic acid strand to the “template” strand. The templatemay be either RNA or DNA, and the complementary strand may also be RNAor DNA. In certain embodiments the complementary strand may comprise allor part of the complementary sequence to the template, or may includemutations so that it is not an exact, complementary strand to thetemplate. Strands that are not exactly complementary to the templatestrand may hybridize specifically to the template strand in detectionassays described here, as well as other assays known in the art, andsuch complementary strands that can be used in detection assays are partof the invention.

[0213] When used in combination with nucleic acid amplificationprocedures, these probes and primers enable the rapid analysis of cell,tissue, or peripheral blood samples. In certain aspects of theinvention, the term “amplification” may refer to any method or techniqueknown in the art or described herein for duplicating or increasing thenumber of copies or amount of a target nucleic acid or its complement.The term “amplicon” refers to the target sequence for amplification, orthat part of a target sequence that is amplified, or the amplificationproducts of the target sequence being amplified. In certain otherembodiments, an “amplicon” may include the sequence of probes or primersused in amplification. This analysis assists in detecting and diagnosinga disease, specifically cardiac, kidney or inflammatory disease, and indetermining optimal treatment courses for individuals at varying stagesof disease progression.

[0214] In light of the present disclosure, one skilled in the art mayselect segments from the identified genes for use in detection,diagnostic, or prognostic methods, vector constructs, antibodyproduction, kits, or any of the embodiments described herein as part ofthe present invention. For example, in certain embodiments the sequencesselected to design probes and primers may include repetitive stretchesof adenine nucleotides (poly-A tails) normally attached at the ends ofthe RNA for the identified differentially expressed gene. In certainother embodiments, probes and primers may be specifically designed tonot include these or other segments from the identified genes, as one ofordinary skill in the art may deem certain segments more suitable foruse in the detection methods disclosed.

[0215] For example, where a genomic sequence is disclosed, one may usesequences that correspond to exon regions of the gene in most cases. Oneskilled in the art may select segments from the published exonsequences, or may assemble them into a reconstructed mRNA sequence thatdoes not contain intronic sequences. Indeed, one skilled in the art mayselect or assemble segments from any of the identified gene sequencesinto other useful forms, such as coding segment reconstructions of MnRNAsequences from published genomic sequences of the identifieddifferentially expressed genes, as part of the present invention. Suchassembled sequences would be useful in designing probes and primers, aswell as providing coding segments for protein translation and fordetection, diagnosis, and prognosis embodiments of the inventiondescribed herein.

[0216] Primers can be designed to amplify transcribed portions of thedifferentially expressed genes of the present invention that wouldinclude any length of nucleotide segment of the transcribed sequences,up to and including the full length of each gene. It is preferred thatthe amplified segments of identified genes be an amplicon of at leastabout 50 to about 500 base pairs in length. It is more preferred thatthe amplified segments of identified genes be an amplicon of at leastabout 100 to about 400 base pairs in length, or no longer in length thanthe amplified segment used to normalize the quantity of message beingamplified in the detection assays described herein. Such assays includeRNA diagnosticing methods, however, differential expression may bedetected by other means, and all such methods would fall within thescope of the present invention. The predicted size of the gene segment,calculated by the location of the primers relative to the transcribedsequence, would be used to determine if the detected amplificationproduct is indeed the gene being amplified. Sequencing the amplified ordetected band that matches the expected size of the amplificationproduct and comparison of the band's sequence to the known or disclosedsequence of the gene would confirm that the correct gene is beingamplified and detected.

[0217] The identified differentially expressed genes may also be used toidentify and isolate full-length gene sequences, including regulatoryelements for gene expression, from genomic human DNA libraries. The cDNAsequences or portions thereof, identified in the present disclosure maybe used as hybridization probes to screen genomic human (or othermammalian) DNA libraries by conventional techniques. Once partialgenomic clones have been identified, “chromosomal walking” may isolatefull-length genes (also called “overlap hybridization”). See Chinault etal, Gene 5:111-26 (1979). Once a partial genomic clone has been isolatedusing a cDNA hybridization probe, nonrepetitive segments at or near theends of the partial genomic clone may be used as hybridization probes infurther genomic library screening, ultimately allowing isolation ofentire gene sequences for the disease, specifically cardiac, kidney orinflammatory disease, state genes of interest. It will be recognizedthat full-length genes may be obtained using small ESTs via technologycurrently available and described in this disclosure (Sambrook et al.,supra; Chinault et al., supra). Sequences identified and isolated bysuch means may be useful in the detection of disease genes using thedetection and diagnostic methods described herein, and are part of theinvention.

[0218] As described before, the identified rat gene may be used as ahybridization probe to screen human or other mammalian cDNA libraries byconventional techniques. Comparison of cloned cDNA sequences with knownhuman or animal cDNA or genomic sequences may be performed usingcomputer programs and databases known in the art.

[0219] The polynucleotides of the present invention are also useful inantisense-mediated gene inhibition, first introduced by Stephenson andZamecnik (Proc. Natl. Acad. Sci. USA 75:285-288 [1978]; see also,Zamecnik et al., Proc. Natl. Acad. Sci. USA 83, 4143-4146 [1986]). Thistechnique is based on the discovery that synthetic DNA fragments caninhibit the transcription and/or translation of selected genes in asequence-specific manner. Since its inception, the technique has foundimportant diagnostic and clinical therapeutic applications in manyfields of oncology, vascular and genetic diseases, and in the treatmentof HIV and other virus infections. To date, two main antisensestrategies have been employed: transfection of cells with antisense cDNAand treatment of cells with antisense oligodeoxynucleotides (ODNs), theuse of ODNs derived from the translation initiation site, e.g., betweenthe −10 and +10 regions of the target gene nucleotide sequence ofinterest being preferred. According to the present invention, moleculescan be designed to reduce or inhibit either normal or, if appropriate,mutant target gene activity, using antisense technology. For furtherdetails see, for example, Wagner, “Gene inhibition using antisenseoligodeoxynucleotides.” Nature 372:333-335 (1992); Tonkinson and Stein,“Antisense oligodeoxynucleotides as clinical therapeutic agents.” CancerInvest. 14:54-65 (1996); Askari and McDonnell,“Antisense-oligonucleotide therapy.” N. Engl. J. Med. 334:316-318(1996); Redekop and Naus, “Transfection with bFGF sense and antisensecDNA resulting in modification of malignant glioma growth.” J.Neurosurg. 82:83-90 (1997); Saleh et al., “Inhibition of growth of C6glioma cells in vivo by expression of antisense vascular endothelialgrowth factor sequence.” Cancer Res. 56:393-401 (1996).

[0220] Oligodeoxynucleotides can be used for the inhibition of genetranscription in the form of triple helix structures. The basecomposition of these oligodeoxynucleotides must be designed to promotetriple helix formation via Hoogsteen base pairing rules, which generallyrequire sizeable stretches of either purines or pyrimidines to bepresent on one strand of a duplex. Nucleotide sequences can bepyrimidine-based, which will result in TAT and CGC+ triplets across thethree associated strands of the resulting triple helix. Thepyrimidine-rich molecules provide base complementarily to a purine-richregion of a single strand of the duplex, in a parallel orientation tothat strand. In addition, nucleic acid molecules can be chosen that arepurine-rich and, for example, contain a stretch of G residues. Thesemolecules form a triple helix with a DNA duplex that is rich in GCpairs, in which the majority of the purine residues are located on asingle strand of the targeted duplex, resulting in GGC triplets acrossthe three strands in the triplex. Alternatively, creating a “switchback”nucleic acid molecule can increase the potential sequences that can betargeted for triple helix formation. Switchback molecules aresynthesized in an alternating 5′-3′, 3′-5′ manner, such that they basepair with first one strand of a duplex and then the other, eliminatingthe necessity for a sizeable stretch of either purines or pyrimidines tobe present on one strand of a duplex.

[0221] The invention also covers the use of ribozymes. Ribozymes areenzymatic RNA molecules capable of catalyzing the specific cleavage ofRNA (Rossi, Current Biology 4:469-71 [1994]). The mechanism of ribozymeaction involves sequence specific hybridization of the ribozyme moleculeto complementary target RNA, followed by an endonucleolytic cleavage.The composition of ribozyme molecules must include one or more sequencescomplementary to the target gene mRNA and must include the well-knowncatalytic sequence responsible for mRNA cleavage. For this sequence, seeU.S. Pat. No. 5,093,246, which is incorporated by reference herein inits entirety. Within the scope of the present invention are engineeredhammerhead motif ribozyme molecules that specifically and efficientlycatalyze endonucleolytic cleavage of RNA sequences encoding target geneproteins.

[0222] Specific ribozyme cleavage sites within any potential RNA targetare initially identified by scanning the molecule of interest forribozyme cleavage sites which include the following sequences, GUA, GWUand GUC. Once identified, short RNA sequences of between 15 and 20ribonucleotides corresponding to the region of the target genecontaining the cleavage site can be evaluated for predicted structuralfeatures, such as secondary structure, that can render theoligonucleotide sequence unsuitable. The suitability of candidatesequences can also be evaluated by testing their accessibility tohybridization with complementary oligonucleotides, using ribonucleaseprotection assays.

[0223] In instances where the antisense, ribozyme, or triple helixmolecules are utilized to reduce or inhibit mutant gene expression, itis possible that the transcription or translation of MRNA produced bynormal alleles is also reduced or inhibited. As a result, theconcentration of normal gene product may be lower than is necessary fora normal phenotype. In such cases, to ensure that substantially normallevels of gene activity are maintained, nucleic acid molecules thatencode and express the polypeptide encoded by the gene targeted, can beintroduced into cells via gene therapy methods, such as those describedbelow. The nucleic acid sequence used in gene therapy is selected suchthat it does not contain sequences susceptible to the antisense,ribozyme, or triple helix treatments utilized. Alternatively, where thetarget gene encodes an extracellular protein, it can be preferable toco-admmister normal target gene protein into the cell or tissue in orderto maintain the requisite level of cellular or tissue target geneactivity.

[0224] The present invention also contemplates the use of “peptidenucleic acids” (PNAs). PNAs have a peptide-like backbone instead of thenormal sugar and phosphate groups of DNA. PNAs may be used to turn onspecific genes, by binding to a promoter region of a gene to initiateRNA transcription. This approach is particularly useful where aparticular disease or disorder is characterized by the underexpressionof a particular gene, or where the increased expression of an identifiedgene has a beneficial effect on the treatment of a disease, inparticular cardiac, kidney or inflammatory disease. Chimeric moleculesof PNA and DNA may also be considered. The DNA portion will allowenzymes attacking DNA-RNA hybrids to cut the RNA part of the complexinto pieces (leading to dissociation of the drug molecule, which canthen be reused), whereas the PNA portion will contribute stability andselectivity.

[0225] As noted before, the polynucleotides of the present invention canalso be used in gene therapy. In gene therapy applications, genes areintroduced into cells in order to achieve in vivo synthesis of atherapeutically effective genetic product, for example for replacementof a defective gene. Gene therapy includes both conventional genetherapy where a lasting effect is achieved by a single treatment, andthe administration of gene therapeutic agents, which involves the onetime or repeated administration of a therapeutically effective DNA orRNA.

[0226] There are a variety of techniques available for introducingnucleic acid into viable cells. The techniques differ depending uponwhether the nucleic acid in transferred into cultured cells in vitro, orin vivo in the cells of the intended host. Techniques suitable for thetransfer of the nucleic acid into mammalian cells in vitro include theuse of liposomes, electroporation, microinjection, cell fusion,DEAE-dextran, the calcium phosphate method, etc. The currently preferredin vivo gene transfer methods include transfection with viral (typicallyretroviral) vectors and viral coat protein-liposome mediatedtransfection (Dzau et al., Trends in Biotechnolozy 11, 205-210 [1993]).In some situations it is desirable to provide the nucleic acid sourcewith an agent that targets the target cells, such as an antibodyspecific for a cell surface membrane protein or the target cells, aligand for a receptor on the target cells, etc. Where liposomes areemployed, proteins which bind to a cell surface membrane proteinassociated with endocytosis may be used for targeting and/or tofacilitate uptake, e.g. capsid proteins or fragments thereof tropic fora particular cell type, antibodies for proteins which undergointernalization in cycling, proteins that target intracellularlocalization and enhance intracellular half-life. For review of genemarking and gene therapy protocols see Anderson et al, Science 256,808-813 (1992).

[0227] The information provided by the present invention can also beused to detect genetic lesions in a differentially expressed gene of thepresent invention, thereby determining if a subject with the lesionedgene is at risk for a disorder characterized by differentially expressedgene expression or polypeptide activity. In preferred embodiments, themethods include detecting, in a biological sample from a subject, thepresence or absence of a genetic lesion characterized by, for example,an alteration affecting the integrity of a gene encoding an polypeptideor the misexpression of the gene. For example, such genetic lesions canbe detected by ascertaining the existence of at least one of: a deletionof one or more nucleotides from a gene; an addition of one or morenucleotides to a gene; a substitution of one or more nucleotides of agene; a chromosomal rearrangement of a gene; an alteration in the levelof a messenger RNA transcript of a gene; aberrant modification of agene, such as of the methylation pattern of the genomic DNA; thepresence of a non-wild type splicing pattern of a messenger RNAtranscript of a gene; a non-wild type level of a gene protein; allelicloss of a gene; and inappropriate post-translational modification of agene protein. As described herein, there are a large number of assaytechniques known in the art that can be used for detecting lesions in agene.

[0228] In certain embodiments, detection of a lesion may involve the useof a probe/primer in, such as anchor PCR or RACE PCR, or, alternatively,in LCR (see, e.g., Landegran et al., Science 241: 1077-80 [1988]; andNakazawa et al., Proc. Natl. Acad. Sci. USA 91: 360-64 [1994]), thelatter of which can be particularly useful for detecting point mutationsin the cardiac gene (see Abravaya et al., Nucleic Acids Res. 23: 675-82[1995]). This method can include the steps of collecting a biologicalsample from a subject, isolating nucleic acid (e.g., genomic, mRNA orboth) from the cells of the sample, contacting the nucleic acid samplewith one or more primers which specifically hybridize to andifferentially expressed gene under conditions such that hybridizationand amplification of the cardiac gene (if present) occurs, and detectingthe presence or absence of an amplification product, or detecting thesize of the amplification product and comparing the length to a controlsample.

[0229] In an alternative embodiment, mutations in a differentiallyexpressed gene from a sample can be identified by alterations inrestriction enzyme cleavage patterns. For example, sample and controlDNA is isolated, amplified (optionally), digested with one or morerestriction endonucleases, and fragment length sizes are determined bygel electrophoresis and compared. Differences in fragment length sizesbetween sample and control DNA indicates mutations in the sample DNA.Moreover, the use of sequence specific ribozymes (see U.S. Pat. No.5,498,531) can be used to score for the presence of specific mutationsby development or loss of a ribozyme cleavage site.

[0230] The arrays of immobilized DNA fragments may also be used forgenetic diagnostics. To illustrate, a microarray containing multipleforms of a mutated gene or genes can be probed with a labeled mixture ofa subject DNA, which will preferentially interact with only one of theimmobilized versions of the gene.

[0231] The detection of this interaction can lead to a medicaldiagnosis. Arrays of immobilized DNA fragments can also be used in DNAprobe diagnostics. For example, the identity of a differentiallyexpressed gene of the present invention can be established unambiguouslyby hybridizing a sample of a subject's DNA to an array comprising knowndifferentially expressed DNA. Other molecules of genetic interest, suchas cDNAs and RNAs can be immobilized on the array or alternately used asthe labeled probe mixture that is applied to the array.

[0232] b. Polypeptides

[0233] The native polypeptides of the present invention, and theirequivalents in other mammalian (e.g. human) species, can be used toidentify interacting proteins and genes encoding such proteins.Interacting proteins and their genes may be part of the signalingpathway in which the differentially expressed genes identified hereinparticipate, and thus are valuable diagnostic and therapeutic candidatesor targets. Among the traditional methods employed areco-immunoprecipitation, cross-linking and co-purification throughgradients or chromatographic columns. Using procedures such as theseallows for the identification of interactive gene products. Onceidentified, an interactive gene product can be used, using standardtechniques, to identify its corresponding interactive gene. For example,at least a portion of the amino acid sequence of the interactive geneproduct can be ascertained using techniques well known to those of skillin the art, such as the Edman degradation technique (see, e.g.,Creighton, Proteins: Structures and Molecular Principles, W. H. Freeman& Co. (New York, N.Y. [1983], pp. 34-49). The amino acid sequenceobtained can be used as a guide for the generation of oligonucleotidemixtures that can be used to screen for interactive gene sequences.Screening can be accomplished, for example, by standard hybridization orPCR techniques. Techniques for the generation of oligonucleotidemixtures and the screening are well known.

[0234] Additionally, methods can be employed which result in thesimultaneous identification of interactive genes that encode the proteininteracting with a protein involved in a disease, specifically 35cardiac, kidney or inflammatory disease. These methods include, forexample, probing expression libraries with a labeled protein known orsuggested to be involved in a disease, using this protein in a mannersimilar to the well known technique of antibody probing of λgtlllibraries.

[0235] A particularly suitable technique for studying protein-proteininteractions is the yeast two-hybrid assay. Many transcriptionalactivators, such as yeast GALA, consist of two physically discretemodular domains, one acting as the DNA-binding domain, while the otherone functioning as the transcription activation domain. The yeasttwo-hybrid system takes advantage of this property, and employs twohybrid proteins, one in which the target protein is fused to theDNA-binding domain of GAL4, and another, in which candidate activatingproteins are fused to the activation domain. The expression of aGALl-calZ reporter gene under control of a GAL4-activated promoterdepends on reconstitution of GAL4 activity via protein-proteininteraction. Colonies containing interacting polypeptides are detectedwith a chromogenic substrate for β-galactosidase. A complete kit(MATCHMAKER™) for identifying protein-protein interactions using theyeast two-hybrid technique is available from Clontech. For furtherdetails see e.g. Fields and Song, Nature (London) 340:245-246 (1989);Chien et al., Proc. Natl. Acad. Sci. USA 88:9578-9582 (1991); andChevray and Nathans, Proc. Natl. Acad. Sci. USA 89:5789-5793 (1992).

[0236] Polypeptides of the present invention may also be used togenerate antibodies, using well-known techniques, some of which havebeen detailed above.

[0237] The polypeptides of the present invention are also useful inassays for identifying lead compounds for therapeutically active agentsfor the treatment of cardiac, kidney or inflammatory diseases. Candidatecompounds include, for example, peptides such as soluble peptides,including Ig-tailed fusion peptides (e.g. immunoadhesins) and members ofrandom peptide libraries (see, e.g., Lam et al., Nature 354:82-84(1991); Houghten et al., Nature 354:84-86 (1991)) and combinatorialchemistry-derived molecular libraries made of D- or L-configurationamino acids; phosphopeptides (e.g., members of random and partiallydegenerate, directed phosphopeptide libraries, see, e.g., Songyang etal, Cell 72:767-78 (1993); antibodies (e.g., polyclonal, monoclonal,humanized, anti-idiotypic, chimeric, and single chain antibodies as wellas Fab, F(ab′)₂, Fab expression library fragments, and epitope-bindingfragments of antibodies); and small organic and inorganic molecules(e.g., molecules obtained from combinatorial and natural productlibraries).

[0238] Such screening assays are preferably amenable to high-throughputscreening of chemical libraries, and are particularly suitable foridentifying small molecule drug candidates. Small molecules, which areusually less than 10 K molecular weight, are desirable as therapeuticssince they are more likely to be permeable to cells, are lesssusceptible to degradation by various cellular mechanisms, and are notas apt to elicit immune response as proteins. Small molecules includebut are not limited to synthetic organic or inorganic compounds, andpeptides. Many pharmaceutical companies have extensive libraries of suchmolecules, which can be conveniently screened by using the assays of thepresent invention. the assays can be performed in a variety of formats,including protein-protein binding assays, biochemical screening assays,immunoassays, cell based assays, etc. Such assay formats are well knownin the art.

[0239] In a preferred embodiment, the screening assays of the presentinvention involve contacting a biological sample obtained from a subjecthaving a disease, specifically cardiac, kidney or inflammatory disease,characterized by the differential expression of a gene identifiedherein, with a candidate compound or agent. The expression of the geneor the activity of the gene product is then determined in the presenceand absence of the test compound or agent. When expression ofdifferentially expressed gene MRNA or polypeptide is greater (preferablystatistically significantly greater) in the presence of the candidatecompound than in its absence, the candidate compound may be identifiedas a stimulator of differentially expressed gene expression.Alternatively, when differentially expressed gene expression is less(preferably statistically significantly less) in the presence of thecandidate compound than in its absence, the candidate compound may beidentified as an inhibitor of differentially expressed gene expression.The level of differentially expressed gene expression in the cells canbe determined by methods described herein for detecting differentiallyexpressed gene mRNA or protein.

[0240] Compounds identified via assays such as those described hereincan be useful, for example, in elaborating the biological function ofthe target gene product, and for treating a cardiac, kidney orinflammatory disease, or ameliorating symptoms of such disease. Ininstances when a disease state or disorder results from a lower overalllevel of target gene expression, target gene product, or target geneproduct activity in a cell involved in the disease, compounds thatinteract with the target gene product can include ones accentuating oramplifying the activity of the bound target gene protein. Such compoundswould bring about an effective increase in the level of target geneactivity, thus treating the disease, disorder or state, or amelioratingits symptoms. Where mutations within the target gene cause aberranttarget gene proteins to be made, which have a deleterious effect thatleads to a disease, compounds that bind target gene protein can beidentified that inhibit the activity of the bound target gene protein.

[0241] 5. Pharmaceutical Compositions

[0242] Pharmaceutical compositions of the present invention can comprisea polynucleotide of the present invention, a product of the genesidentified herein, or other therapeutically active compounds, includingorganic small molecules, peptides, polypeptides, antibodies etc.identified with the aid of the differentially expressed genes identifiedherein.

[0243] Suitable forms, in part, depend upon the use or the route ofentry, for example oral, transdermal, inhalation, or by injection. Suchforms should allow the agent or composition to reach a target cellwhether the target cell is present in a multicellular host or inculture. For example, pharmacological agents or compositions injectedinto the blood stream should be soluble. Other factors are known in theart, and include considerations such as toxicity and forms that preventthe agent or composition from exerting its effect.

[0244] The active ingredient, when appropriate, can also be formulatedas pharmaceutically acceptable salts (e.g., acid addition salts) and/orcomplexes. Pharmaceutically acceptable salts are non-toxic at theconcentration at which they are administered. Pharmaceuticallyacceptable salts include acid addition salts such as those containingsulfate, hydrochloride, phosphate, sulfonate, sulfamate, sulfate,acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate, cyclolexylsulfonate,cyclohexylsulfamate and quinate. Pharmaceutically acceptable salts canbe obtained from acids such as hydrochloric acid, sulfuric acid,phosphoric acid, sulfonic acid, sulfamic acid, acetic acid, citric acid,lactic acid, tartaric acid, malonic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,cyclohexylsulfonic acid, cyclohexylsulfamic acid, and quinic acid. Suchsalts may be prepared by, for example, reacting the free acid or baseforms of the product with one or more equivalents of the appropriatebase or acid in a solvent or medium in which the salt is insoluble, orin a solvent such as water which is then removed in vacuo or byfreeze-drying or by exchanging the ions of an existing salt for anotherion on a suitable ion exchange resin.

[0245] Carriers or excipients can also be used to facilitateadministration of the compound. Examples of carriers and excipientsinclude calcium carbonate, calcium phosphate, various sugars such aslactose, glucose, or sucrose, or types of starch, cellulose derivatives,gelatin, vegetable oils, polyethylene glycols and physiologicallycompatible solvents. The compositions or pharmaceutical composition canbe administered by different routes including, but not limited to,intravenous, intra-arterial, intraperitoneal, intrapericardial,intracoronary, subcutaneous, and intramuscular, oral, topical, ortransmucosal.

[0246] The desired isotonicity of the compositions can be accomplishedusing sodium chloride or other pharmaceutically acceptable agents suchas dextrose, boric acid, sodium tartrate, propylene glycol, polyols(such as mannitol and sorbitol), or other inorganic or organic solutes.

[0247] Pharmaceutical compositions can be formulated for a variety ofmodes of administration, including systemic and topical or localizedadministration. Techniques and formulations generally may be found inRemington's Pharmaceutical Sciences, 18^(th) Edition, Mack PublishingCo., Easton, Pa. 1990. See also, Wang and Hanson “ParenteralFormulations of Proteins and Peptides: Stability and Stabilizers”,Journal of Parenteral Science and Technology, Technical Report No. 10,Supp. 42-2S (1988). A suitable administration format can best bedetermined by a medical practitioner for each patient individually.

[0248] For systemic administration, injection is preferred, e.g.,intramuscular, intravenous, intra-arterial, intracoronary,intrapericardial, intraperitoneal, subcutaneous, intrathecal, orintracerebrovascular. For injection, the compounds of the invention areformulated in liquid solutions, preferably in physiologically compatiblebuffers such as Hank's solution or Ringer's solution. Alternatively, thecompounds of the invention are formulated in one or more excipients(e.g., propylene glycol) that are generally accepted as safe as definedby USP standards. They can, for example, be suspended in an inert oil,suitably a vegetable oil such as sesame, peanut, olive oil, or otheracceptable carrier. Preferably, they are suspended in an aqueouscarrier, for example, in an isotonic buffer solution at pH of about 5.6to 7.4. These compositions can be sterilized by conventionalsterilization techniques, or can be sterile filtered. The compositionscan contain pharmaceutically acceptable auxiliary substances as requiredto approximate physiological conditions, such as pH buffering agents.Usefuil buffers include for example, sodium acetate/acetic acid buffers.A form of repository or “depot” slow release preparation can be used sothat therapeutically effective amounts of the preparation are deliveredinto the bloodstream over many hours or days following transdermalinjection or delivery. In addition, the compounds can be formulated insolid form and redissolved or suspended immediately prior to use.Lyophilized forms are also included.

[0249] Alternatively, certain compounds identified in accordance withthe present invention can be administered orally. For oraladministration, the compounds are formulated into conventional oraldosage forms such as capsules, tablets and tonics.

[0250] Systemic administration can also be by transmucosal ortransdernal. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, bile salts and fusidic acidderivatives. In addition, detergents can be used to facilitatepermeation. Transmucosal administration can be, for example, throughnasal sprays or using suppositories.

[0251] For administration by inhalation, usually inhalable dry powercompositions or aerosol compositions are used, where the size of theparticles or droplets is selected to ensure deposition of the activeingredient in the desired part of the respiratory tract, e.g. throat,upper respiratory tract or lungs. Inhalable compositions and devices fortheir administration are well known in the art. For example, devices forthe delivery of aerosol medications for inspiration are known. One suchdevice is a metered dose inhaler that delivers the same dosage ofmedication to the patient upon each actuation of the device. Metereddose inhalers typically include a canister containing a reservoir ofmedication and propellant under pressure and a fixed volume metered dosechamber. The canister is inserted into a receptacle in a body or basehaving a mouthpiece or nosepiece for delivering medication to thepatient. The patient uses the device by manually pressing the canisterinto the body to close a filling valve and capture a metered dose ofmedication inside the chamber and to open a release valve which releasesthe captured, fixed volume of medication in the dose chamber to theatmosphere as an aerosol mist. Simultaneously, the patient inhalesthrough the mouthpiece to entrain the mist into the airway. The patientthen releases the canister so that the release valve closes and thefilling valve opens to refill the dose chamber for the nextadministration of medication. See, for example, U.S. Pat. No. 4,896,832and a product available from 3M Healthcare known as Aerosol SheathedActuator and Cap.

[0252] Another device is the breath actuated metered dose inhaler thatoperates to provide automatically a metered dose in response to thepatient's inspiratory effort. One style of breath actuated devicereleases a dose when the inspiratory effort moves a mechanical lever totrigger the release valve. Another style releases the dose when thedetected flow rises above a preset threshold, as detected by a hot wireanemometer. See, for example, U.S. Pat. Nos. 3,187,748; 3,565,070;3,814,297; 3,826,413; 4,592,348; 4,648,393; 4,803,978.

[0253] Devices also exist to deliver dry powdered drugs to the patient'sairways (see, e.g. U.S. Pat. No. 4,527,769) and to deliver an aerosol byheating a solid aerosol precursor material (see, e.g. U.S. Pat. No.4,922,901). These devices typically operate to deliver the drug duringthe early stages of the patient's inspiration by relying on thepatient's inspiratory flow to draw the drug out of the reservoir intothe airway or to actuate a heating element to vaporize the solid aerosolprecursor.

[0254] Devices for controlling particle size of an aerosol are alsoknown, see, for example, U.S. Pat. Nos. 4,790,305; 4,926,852; 4,677,975;and 3,658,059.

[0255] For topical administration, the compounds of the invention areformulated into ointments, salves, gels, or creams, as is generallyknown in the art.

[0256] If desired, solutions of the above compositions can be thickenedwith a thickening agent such as methyl cellulose. They can be preparedin emulsified form, either water in oil or oil in water. Any of a widevariety of pharmaceutically acceptable emulsifying agents can beemployed including, for example, acacia powder, a non-ionic surfactant(such as a Tween), or an ionic surfactant (such as alkali polyetheralcohol sulfates or sulfonates, e.g., a Triton).

[0257] Compositions useful in the invention are prepared by mixing theingredients following generally accepted procedures. For example, theselected components can be mixed simply in a blender or other standarddevice to produce a concentrated mixture which can then be adjusted tothe final concentration and viscosity by the addition of water orthickening agent and possibly a buffer to control pH or an additionalsolute to control tonicity.

[0258] The amounts of various compounds for use in the methods of theinvention to be administered can be determined by standard procedures.Generally, a therapeutically effective amount is between about 100 mg/kgand 10⁻¹² mg/kg depending on the age and size of the patient, and thedisease or disorder associated with the patient. Generally, it is anamount between about 0.05 and 50 mg/kg of the individual to be treated.The determination of the actual dose is well within the skill of anordinary physician.

[0259] The invention is further illustrated in the followingnon-limiting examples.

EXAMPLES Example 1

[0260] Identification of Differentially Expressed Rat Genes Referred toby Clone ID Number

[0261] 1. In vivo model of myocardial infarction

[0262] Genes P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3),P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ IDNO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08(SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19),P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ IDNO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00223_F07(SEQ ID NO:31), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34),P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ IDNO:40), P00239_C11 (SEQ ID NO:42), P00240_B04 (SEQ ID NO:44), P00240_E05(SEQ ID NO:45), P00241_E12 (SEQ ID NO:47), P00245_D06 (SEQ ID NO:48),P00246_D12 (SEQ ID NO:49), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ IDNO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10(SEQ ID NO:58), P00263_G06 (SEQ ID NO:60), P00267_F08 (SEQ ID NO:61),P00269_H08 (SEQ ID NO:62), P00312_C04 (SEQ ID NO:64), P00324_H02 (SEQ IDNO:65), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00634_G11(SEQ ID NO:70), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73),and P00697_C03 (SEQ ID NO:75), were identified by analysis of leftventricular heart tissue obtained from an in vivo model of leftventricle myocardial infarction (MI) (Pfeffer et al, Circ. Res. 57:84-95[1985]). Specifically, male Sprague-Dawley rats at age 7-10 weeks wereanesthetized with ketamine (80 mg/kg IP) and xylazine (10 mg/kg IP). Thethorax and abdomen was shaved, after which the areas were scrubbed withprovidone-iodine and 70% isopropyl alcohol a minimum of three times,beginning at the incision line and continuing in a circular motionproceeding toward the periphery. The rats were intubated and placed on arespirator with room air at a rate of 55 breaths/min. A left thoracotomywas performed between the fourth and fifth ribs, after which the heartwas exteriorized and the left anterior descending coronary artery (LAD)ligated with silk suture. The same surgical procedure was employed forsham-operated rats, however, the suture was passed through the leftventricular wall and the LAD was not occluded.

[0263] Following the surgical procedure, negative pressure in thethoracic was quickly reestablished and the wound closed with apurse-string suture using 3-0 non-absorbable suture material.Butorphanoll (0.1 mg/kg. SQ) was provided post surgery as a prophylacticanalgesic. The rats were extubated when they recovered their gag reflexand allowed recovering in a warming chamber. Seventy-five percent of therats had large infarcts on their left ventricle free walls andperioperative mortality rate is about 50%, which is comparable to thepublished data.

[0264] Tissue was collected 2 week, 4 week, 8 week, 12 week and 16 weekpost-surgery. Blood was collected the day before surgery and the daybefore sacrifice for measurement of plasma atrial natriuretic peptide(ANP) level. On the day of necropsy, each heart was divided transverselyinto two halves so that the infarcted area is bisected. One half of theheart was used for histological evaluation, and the other for mRNAmicroarray analysis.

[0265] 2. In vivo Model of Septum Myocardial Infarction

[0266] Septum tissue was obtained from diseased rat hearts obtainedthrough the left ventricle rat MI model of Pfeffer et al., as describedabove. Poly A+ MRNA was prepared from each of these septums forassessment of differentially expressed genes in the disease state, usingmicroarray analysis in a preferred embodiment.

[0267] 3. Preparation of normalized CDNA libraries

[0268] Poly A+ mRNA was prepared from each of the animals, forassessment of differentially expressed genes in the disease state, usingmicroarray analysis. Total RNA was isolated from homogenized tissue byacid phenol extraction (Chomczynski and Sacchi, Anal. Biochem.162(1):156-9 [1987]). Poly A+ MRNA was selected from total RNA by oligodT hybridization utilizing a polyA Spin mRNA Isolation Kit (New EnglandBioLabs, Beverly, Mass.) according to manufacturers' protocols. Adirectionally cloned cDNA library was first generated by conventionalmethods. Briefly, double stranded cDNA was generated by priming firststrand synthesis for reverse transcription using oligo dT primers whichcontain a Not I restriction site. After second strand synthesis, Xba Iadapters were added to the 5′ end of the cDNA, and the cDNA size wasselected for >500 bp and ligated into the corresponding restrictionsites of phagemid vector pCR2.1 (Invitrogen, San Diego Calif.).

[0269] From the total cDNA library, a normalized library was generatedas detailed elsewhere (see, e.g. Bonaldo et al, Genome Res. 6(9):791-806[1996]) and described here briefly. Phagemid vector pCR2.1 contains anF1 origin of replication. Thus, the cDNA library can be propagated assingle stranded phage with an appropriate helper virus. Single stranded,circular DNA was extracted from the phage library and served as “tester”DNA in the hybridization step of normalization. The other component ofthe hybridization, “driver” DNA, was generated from the library by PCRamplification using a set of the following primers specific for theregion of the vector, which flanks the cloned inserts:5′CGTATGTTGTGTGGAATTGTGAGCG (SEQ ID NO:77) 5′GATGTGCTGCAAGGCGATTAAGTTG(SEQ ID NO:78)

[0270] Purified tester DNA (50 ng) and driver DNA (0.5 μg) were combinedin 120 mM NaCl, 50% formamide, 10 mM Tris (pH 8.0), 5 mM EDTA, and 1%SDS. A set of oligonucleotides (10 μg each), corresponding to polylinkersequence (same strand as tester DNA) which is present in the PCRproduct, was included in the hybridization reaction to block annealingof vector-specific sequences which are in common between tester anddriver DNA. The oligonucleotide sequences were as follows:5′GCCGCCAGTGTGCTGGAATTCGGCTAGC (SEQ ID NO: 79)5′CGAATTCTGCAGATATCCATCACACTGG (SEQ ID NO: 80)5′CTAGAGGGCCCAATTCGCCCTATAG (SEQ ID NO: 81) 5′TGAGTCGTATTACAATTCACTGGCC(SEQ ID NO: 82) 5′GCTCGGATCCACTAGTAACG (SEQ ID NO: 83)5′TTTTTTTTTTTTTTTTTT (SEQ ID NO: 84)

[0271] The reaction mixture, under oil, was heated 3 min. at 80° C., andhybridization performed at 30° C. for 24 hr (calculated C_(o)t˜5).Single stranded circles were purified from the reaction mixture byhydroxylapatite (HAP) chromatography, converted to double strand DNA,and electroporated into bacteria to yield a normalized cDNA libraryrepresentative of genes expressed in the left ventricle of rat. Toevaluate the effectiveness of the normalization protocol, the frequencyof a few clones (ANP, BNP, actin, and myosin) was assessed in both inthe starting library and the normalized library. The frequency ofabundant cDNAs (actin and myosin) was reduced and roughly equivalent torarer cDNA clones (ANP and BNP). Clone frequency in the two librarieswas determined with standard screening techniques by immobilizingcolonies onto nylon membranes and hybridizing with radiolabeled DNAprobes.

[0272] Certain genes, unexpressed in a normal tissue and turned on indiseased tissue, may be absent from the normalized cDNA librarygenerated from normal tissue. To obtain disease-specific clones toinclude on the microarray, one can repeat the normalization strategyusing diseased tissue obtained from the appropriate disease model.However, since most genes are expressed commonly between normal anddiseased tissue, microarraying normalized libraries from diseased andnormal tissue may introduce significant redundancy, a subtracted librarycan be made using protocols similar to those used to generate normalizedlibraries. Again, the method of Bonaldo et al., supra, as described herebriefly, is used.

[0273] To make a subtracted library, a total cDNA library is generatedfrom the tissue obtained from the disease model (e.g., left ventricletaken from the MI Model). The cDNA library is directionally cloned inpCR2.1 vector and single stranded tester DNA derived as described abovefor library normalization. The driver DNA is generated by PCRamplification of cloned inserts from the total cDNA library preparedfrom the left ventricle of normal rat. Hybridization occurs betweensequences, which are in common to normal and diseased hearts. For thissubtracted library, the reaction is driven more thoroughly (calculatedC_(ot)˜27) than normalization by using more driver (1.5 μg vs. 0.5 μg)and longer hybridization time (48 hr vs. 24 hr). Purification ofnonhybridized, single stranded circles by HAP chromatography, conversionto double strand DNA, and electroporation into bacteria yields asubtracted cDNA library enriched for genes which are expressed indiseased rat hearts. To test that the library is truly subtracted,colony hybridization is performed with probes for ANP, BNP, actin, andmyosin. The subtracted library has a high frequency of ANP and BNPclones since they are elevated significantly in the hypertrophic ratheart. Actin and myosin clones are absent since they are expressedequally in normal and diseased left ventricle.

[0274] 4. Microarray Analysis

[0275] High quality DNA is important for the microarray printingprocess. A microtiter plate protocol for PCR amplification of DNA andits subsequent purification was established that provides acceptablequality and quantity of DNA for printing on microarrays. Specifically,the following PCR probes were synthesized that amplify insert DNA fromthe vector pCR2.1 that was used for library construction.:5′CGTATGTTGTGTGGAATTGTGAGCG (SEQ ID NO: 85) 5′GATGTGCTGCAAGGCGATTAAGTTG(SEQ ID NO: 86)

[0276] After 30 cycles of amplification each PCR product was passed overa gel filtration column to remove unincorporated primers and salts. Tomaintain robustness, the columns were packed in 96-well filter platesand liquid handling was performed using a robotic liquid handler (Biomek2000, Beckman).

[0277] To test the quality of DNA prepared by this PCR method, 96purified samples from a single microtiter plate were produced as amicroarray. Using the robotic liquid handler, 85 pl of PCR reactionmixture was aliquoted into each well of a thin walled, 0.2 ml 96-wellplate. The reaction mixture contained 0.2 mM each dNTP, 1.25 units ofTaq polymerase, and 1× Taq buffer (Boehringer Mannheim). Primers, 1 μmeach, are from vector regions, which flank the cloning site of pCR2.1and include a 5′ primary amine with a 6-carbon linker to facilitateattachment of DNA product to the glass surface of the microarray chip.1.0 μl of bacterial culture of individual cDNA clones was added to eachwell. PCR conditions were: 2 min., 95° C. to denature, then 30 cycles of95° C., 30 sec./65° C., 40 sec./72° C., 1 min. 30 sec., and a finalextension of 72° C., 5 min. using a MJResearch PTC 100 thermocycler.

[0278] PCR products were purified by gel filtration over Sephacryl 400(Sigma). Briefly, 400 μl of pre-swollen Sephacryl 400 was loaded intoeach well of a 96-well filter plate (PallBiosupport) and spun into acollection plate at 800 g for 1 min. Wells were washed 5 times with 0.2×SSC. PCR reaction mixtures were loaded onto the column and purified DNA(flow-through) was collected at 800 g for 1 min. Samples were dried downat 50° C. overnight and arrayed.

[0279] Fluorescent probe pairs were synthesized by reverse transcriptionof poly A+ RNA using, separately, Cy3 dCTP and Cy5 dCTP (Amersham). In16.5 μl, 1 μg poly A+ RNA and 2 μg of oligo dT 21 mer, were denatured at65° C., 5 min. and annealed at 25° C., 10 min. Reverse transcription wasperformed for 2 hours at 37° C. with Superscript RT (Life Technologies,Gaithersburg, Md.) in 1× buffer, 10 units RNase block, 500 μM eachdATP/dGTP/dTTP, 280 μM dCTP, 40 μM Cy5 or Cy3 dCTP, and 200 units RT.RNA is degraded in 0.1 M NaOH, 65° C. for 10 min. Labeled cDNA waspurified by successive filtration with Chroma Spin 30 spin columns(Clontech) following manufacturer's instructions. Samples were dried atroom temperature in the dark using a covered Speed-Vac. Probes wereapplied to the test chip for hybridization and the data collectedessentially as described in Schena et al., cited above The intensity ofhybridization signal at each element reflected the level of expressionof the mRNA for each gene in the rat ventricle. Digitized signal datawas stored and prepared for analysis.

[0280] A series of control DNA elements were included on each chip toensure consistency in labeling and hybridization between experiments andto aid in balancing the signal when two fluorescence channels are used.For each element hybridized with dual labeled probes, absolute andrelative intensity of signal was determined. The results from these andother experiments indicate that these methods for production of templateDNA and labeled cDNA probes are suitable for generating high qualitymicroarrays within a preferred embodiment of the methods of the presentinvention. The evaluation of tens of thousands of genes for expressiongenerates a large amount of data that can be manipulated by commerciallyavailable software packages that facilitate handling this type andquantity of data. The expression data can be stored, analyzed, andsorted from each experiment using this software. In addition, expressionof each clone can be tracked from experiment to experiment using knownmethodologies.

[0281] The novel secreted factor of the present invention was identifiedfrom expression data from the following experiments: A 10,000 clonemicroarray (10 K) from a normalized normal rat left ventricle (LV) cDNAlibrary was probed in duplicate. A 3,000 clone array, which includeddifferentially expressed clones from the 1 OK library, was also probedin duplicate. Included on the microarray with the unidentified geneswere a set of known clones. These known clones were included becausethey represent genes of particular interest and help evaluate thesensitivity of the microarray methodology. Indeed, any genes ofparticular interest may be included on such microarrays. By way ofexample, ANP, BNP, endothelin, β-myosin heavy chain, and oc-actin aregenes that change expression levels in the LVH model, and thus theyserve as useful positive controls in the in vivo model exemplifiedherein.

[0282] The intensity of hybridization signal at each element of themicroarray reflected the level of expression of the MRNA for each gene.For each element hybridized with dual labeled probes, absolute andrelative intensity of signal was determined, which translates into therelative expression levels of the subject genes. The numeric dataobtained reflect the relative expression level of the gene in thedisease state as compared to the expression level of the gene in thenormal, or non-disease state. Positive numbers are indicative of genesexpressed at higher levels in the diseased tissue relative to normaltissue, and negative values are indicative of lower expression indisease. Data are the average values from multiple experiments performedwith separate DNA arrays (n=4 for MI left ventricle and septum). Arrayprobes were generated from RNA pooled from multiple animals (n=4 forMI).

[0283] The data also reflect expression levels of genes in certaindisease models over various time points. For example, gene expression inthe myocardial infarction model was compared at 2, 4, 8, 12, and 16weeks for the representative genes in the disease state versus thenormal state. Indeed, such experimentation provides valuable dataregarding the temporal relationship of gene expression levels in diseasestates and provides important insights regarding the treatment,diagnosis, and modulation of differentially expressed disease stategenes, as discussed in detail infra.

[0284] One to two percent of the clones assayed on microarrays werefound to be differentially expressed. Secondary chips may be used formore extensive hybridizations, including examination of individualanimals, and more thorough evaluation of time points. In a preferredembodiment, clones that reproducibly scored in microarray analysis to beat least about 1.8-fold elevated or decreased were microarrayed onseparate secondary chips and their expression levels determined. It isunderstood, however, that differentially expressed genes exhibiting lessthan about a two-fold change in expression, e.g., less than one,one-half, or one-quarter, or greater than about a two-fold change inexpression, e.g., greater than three, five, ten, twenty, onehundred-fold, or one thousand-fold, are within the scope of the presentinvention.

[0285] 5. Microarray Results

[0286] Using the foregoing protocols, it was found that in the MI model,the expression level of the gene corresponding to the clones weredifferentially expressed in heart and kidney. This differentialexpression suggests the possible involvement of these genes in thedevelopment and/or progress of MI. The results are summarized in FIG.44.

[0287] 6. Sequence Analysis

[0288] The differentially expressed partial and full-length clonesP00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ IDNO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05(SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15),P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ IDNO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05(SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00223_F07 (SEQ ID NO:31),P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ IDNO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11(SEQ ID NO:42), P00240_B04 (SEQ ID NO:44), P00240_E05 (SEQ ID NO:45),P00241_E12 (SEQ ID NO:47), P00245_D06 (SEQ ID NO:48), P00246_D12 (SEQ IDNO:49), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09(SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58),P00263_G06 (SEQ ID NO:60), P00267_F08 (SEQ ID NO:61), P00269_H08 (SEQ IDNO:62), P00312_C04 (SEQ ID NO:64), P00324_H02 (SEQ ID NO:65), P00628_H02(SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00634_G11 (SEQ ID NO:70),P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), and P00697_C03(SEQ ID NO:75) were sequenced (SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15,17, 19, 21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40, 42, 44, 45,47, 48,49, 50, 52, 54, 56, 58, 59, 60, 61, 62, 64, 65, 66, 68, 70, 71, 73, and75), and the deduced amnino acid sequence was determined (SEQ ID NOs: 2,4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41,43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, and 76). FIGS. 1-43 showthe deduced amnino acid sequence of the polypeptide encoded by theclones as well as the nucleotide sequences.

[0289] The nucleotide sequences of the clones were compared withsequences in the public GenBank, EMBL, DDBJ, PDB and GENSEQ databases.The search was performed using the BLASTN 2.0.8 program with defaultparameters. Gap penalties: existence: 5; extension: 2. The searchrevealed no significant homology with sequences present in the searcheddatabases.

[0290] 7. Northern blot analysis

[0291] Northern blot analysis suggested that the clones aredifferentially expressed (see FIG. 44).

Example 2

[0292] Identification of the Human Homologue of Rat Clone

[0293] The isolated differentially expressed rat gene sequence can belabeled and used to screen a cDNA library constructed from mRNA obtainedfrom an organism of interest. Hybridization conditions will be of alower stringency when the cDNA library was derived from an organismdifferent from the type of organism from which the labeled sequence wasderived. Alternatively, the labeled fragment can be used to screen agenomic library derived from the organism of interest, again, usingappropriately stringent conditions. Such low stringency conditions willbe well known to those of skill in the art, and will vary predictablydepending on the specific organisms from which the library and thelabeled sequences are derived. For guidance regarding such conditionssee, Sambrook et al., supra, and Ausubel et al., supra.

[0294] PCR technology can also be utilized to isolate full-length humancDNA sequences. For example, RNA can be isolated, following standardprocedures, from an appropriate human cellular or tissue source. Areverse transcription reaction can be performed on the RNA using anoligonucleotide primer specific for the most 5′ end of the amplifiedfragment for the priming of first strand synthesis. The resultingRNA/DNA hybrid can then be “tailed” with guanines using a standardterminal transferase reaction, the hybrid can be digested with RNase H,and second strand synthesis can then be primed with a poly-C primer.Thus, cDNA sequences upstream of the amplified fragment can easily beisolated. For a review of cloning strategies that can be used, see,e.g., Sambrook et al., supra, and Ausubel et aL, supra.

[0295] Alternatively, the human homologue can be isolated using theCloneCapture cDNA selection Kit (Clontech, Palo Alto, Calif.): aRecA-based system for the rapid enrichment and isolation of cDNA clonesof interest without library screening.

Example 3

[0296] Expression of the Clones in E. coli

[0297] The P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3),P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ IDNO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08(SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19),P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ IDNO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01(SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36),P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ IDNO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04(SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56),P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ IDNO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12(SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75) DNA is initially amplifiedusing selected PCR primers. The primers should contain restrictionenzyme sites that correspond to the restriction enzyme sites on theselected expression vector. A variety of expression vectors may beemployed. An example of a suitable vector is pBR322 (derived from E.coli; see Bolivar et al., Gene, 2:95 [1977]) which contains genes forampicillin and tetracycline resistance, or a pBR322-based vector. Other,commercially available vectors include various pUC vectors andBluescript M13. The vector is digested with restriction enzyme anddephosphorylated. The PCR amplified sequences are then ligated into thevector. The vector will preferably include sequences that encode anantibiotic resistance gene, a promoter, such as a T7 or tryptophan (trp)promoter, a polyhis leader (including the first six STII codons, polyhissequence, and enterokinase cleavage site), the P00184_D11 (SEQ ID NO:1),P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ IDNO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:1l), P00194_H10(SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17),P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ IDNO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03(SEQ ID NO:29), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34),P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ IDNO:40), P00239_C11 (SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04(SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54),P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ IDNO:62), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11(SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75)coding region, lambda transcriptional terminator, and an argu gene.

[0298] The ligation mixture is then used to transform a selected E. colistrain using the methods described in Sambrook et al., supra.Transformants are identified by their ability to grow on LB plates andantibiotic resistant colonies are then selected. Plasmid DNA can beisolated and confrmned by restriction analysis and DNA sequencing.

[0299] Selected clones can be grown overnight in liquid culture mediumsuch as LB broth supplemented with antibiotics. The overnight culturemay subsequently be used to inoculate a larger scale culture. The cellsare then grown to a desired optical density, during which the expressionpromoter is turned on.

[0300] After culturing the cells for several more hours, the cells canbe harvested by centrifugation. The cell pellet obtained by thecentrifugation can be solubilized using various agents known in the art,and the solubilized protein can then be purified using a metal chelatingcolumn under conditions that allow tight binding of the poly-his taggedprotein.

Example 4

[0301] Expression of the Clones in Yeast

[0302] A yeast expression vector is constructed either for intracellularproduction or secretion of the protein encoded by P00184_D11 (SEQ IDNO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01(SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11),P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ IDNO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02(SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27),P00222_G03 (SEQ ID NO:29), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ IDNO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08(SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05 (SEQ ID NO:45),P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ IDNO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00269_H08(SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68),P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), and P00697_C03(SEQ ID NO:75), using an appropriate yeast promoter, such the promoterof 3-phosphoglycerate kinase, or the promoter regions for alcoholoxidase 1 (AOX1, particularly preferred for expression in Pichia),alcohol dehydrogenase 2, or isocytochrome C. For secretion, theP00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ IDNO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05(SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15),P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ IDNO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05(SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01 (SEQ ID NO:32),P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ IDNO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05(SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52),P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ IDNO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08(SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73),and P00697_C03 (SEQ ID NO:75) coding sequence is linked, at its 5′-end,to a mammalian or yeast signal (secretory leader) sequence, such as ayeast alpha-factor or invertase secretory signal. Alternatively, acommercially available yeast expression system is used that can bepurchased, for example, from Clontech Laboratories, Inc. (Palo Alto,Calif., e.g. pYEX 4T family of vectors for Saccharomyces cerevisiae),Invitrogen (Carlsbad, Calif., e.g. pPICZ series Easy Select PichiaExpression Kit) or Stratagene (La Jolla, Calif., e.g. ESP™ Yeast ProteinExpression and Purification System for S. pombe and pESC vectors for S.cerevisiae).

[0303] Yeast cells, such as S. cerevisiae AB110 strain, or P. pastorisGS115 (NRRL Y-15851); GS190 (NRRL Y-18014) or PPF1 (NRRL Y-18017) arethen transformed by known techniques, e.g. by the polyethylene glycolmethod (Hinnen, Proc. Natl. Acad, Sci. USA 75:1929 [1978]).

[0304] The recombinant protein is subsequently isolated and purified byremoving the yeast cells from the fermentation medium by centrifugationand then concentrating the medium using selected cartridge filters. Theconcentrate containing the expressed protein may be further purifiedusing selected column chromatography resins.

Example 5

[0305] Expression of the Clones in Mammalian Host Cells

[0306] The P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3),P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ IDNO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08(SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19),P00209_F06 (SEQ ID NO:21), P06219_D02 (SEQ ID NO:23), P00219_F06 (SEQ IDNO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01(SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36),P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ IDNO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04(SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56),P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ IDNO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12(SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75) genes are subjected to PCRusing primers containing suitable restriction enzyme cleavage sites toallow ligation into a mammalian expression vector such as pCEP4(Invitrogen). To facilitate the eventual recovery of the expressedprotein, it is advisable to use the 3′ PCR primer to extend the openreading frame of the cloned gene to include an affinity purification tagsuch as poly-His (E. Hochuli et al 1987, J. Chrom. 411, 177-184) orcalmodulin binding peptide (Hathaway et al, J Biol. Chem. 1981,256(15):8183-9). Recovery of the PCR fragment may be followed by itscleavage at the new flanking restriction sites and ligation into asimilarly cleaved pCEP4 preparation. Transformation of bacteria andpreparation of plasmids from transformants is followed by verificationof the plasmid structure by restriction analysis.

[0307] Expression of the P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ IDNO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01(SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13),P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ IDNO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06(SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29),P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ IDNO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11(SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50),P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ IDNO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02(SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71),P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75) genes can beaccomplished by transient expression in 293 human embryonic kidneycells. For use of vectors such as pCEP4 having the EBV viral origin ofreplication, 293EBNA cells that are permissive for replication can beused. Transfection is accomplished using a lipid transfection reagentsuch as Lipofectamine Plus (Life Technologies, Rockville, Md.).Endotoxin-free plasmid DNA (100 μg) is added to 200 μl PLUS reagent and10 ml DMEM-21 serum free media to give Mix A. This is incubated at roomtemperature for 15 minutes. Mix B is prepared from 400 μl Lipofectamineand 10 ml serum-free DMEM-21. The two mixes are then combined andincubated at room temperature for another 15 minutes. An 850 cm² rollerbottle containing the cells to be transfected at 70% confluence isrinsed with serum-free media and 100 ml of serum-free DMEM-2 with 15 mMHEPES pH 7.3 and the DNA-lipid transfection mixture is then added. Thecells are then placed in a roller unit at 37 C for 4 hours after whichthe volume of media is doubled by addition of DMEM-2 with 15 mM HEPES pH7.3, 5% FBS and the bottle returned to roller unit overnight. Collectconditioned media every 2-3 days for 2-3 collections.

Example 6 Expression of the Clones in Baculovirus-Infected Insect Cells

[0308] Baculovirus-based expression is performed using one of thecoummercially available baculovirus expression systems such as, forexample, from Bac-N-Blue™ (Invitrogen), BacPAK™ Baculovirus ExpressionSystem (Clontech), BAC-TO-BAC™ (Life Technologies), or Bac VectorSystem™ (Novagen). Viral infection of insect cells (e.g. Spodopterafrugiperda (“Sf9”) cells (ATCC CRL 1711)) and protein expression andpurification are performed following manufacturers' instructions, or asdescribed by O'Reilley et al., Baculovirus expression vectors: ALaboratorv Manual, Oxford: Oxford University Press (1994). Optionally,the coding region of the P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ IDNO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01(SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13),P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ IDNO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06(SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29),P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ IDNO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11(SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50),P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ IDNO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02(SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71),P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75) sequence isfused upstream of an epitope tag contained within a baculovirusexpression vector, such as a poly-His tag or an imumunoglobulin (Ig) tag(like Fc regions of an IgG). The poly-His or Ig tag aids proteinpurification.

Example 7

[0309] Preparation of antibodies that bind the polypeptide encoded byP00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ IDNO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05(SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15),P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ IDNO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05(SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01 (SEQ ID NO:32),P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ IDNO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05(SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52),P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ IDNO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08(SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73),and P00697_C03 (SEQ ID NO:75)

[0310] This example illustrates preparation of monoclonal antibodiesthat specifically bind the polypeptide encoded by P00184_D11 (SEQ IDNO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01(SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11),P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ IDNO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02(SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27),P00222_G03 (SEQ ID NO:29), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ IDNO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08(SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05 (SEQ ID NO:45),P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ IDNO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00269_H08(SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68),P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), and P00697_C03(SEQ ID NO:75).

[0311] Techniques for producing the monoclonal antibodies are known inthe art and are described, for instance, in Goding, supra. Theinummunogen may, for example, be purified protein encoded by the cloneor recombinant host cells expressing P00184_D11 (SEQ ID NO:1),P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ IDNO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10(SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17),P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ IDNO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03(SEQ ID NO:29), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34),P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ IDNO:40), P00239_C11 (SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04(SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54),P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ IDNO:62), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11(SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ IDNO:75). Mice, such as Balb/c, are immunized with the immunogenemulsified in a selected adjuvant, for example Freund's adjuvant, andinjected subcutaneously or intraperitoneally in an amount from 1-100micrograms. Approximately 10 to 12 days later, the inununized mice areboosted with additional imnunogen emulsified in the selected adjuvant.Thereafter, for several weeks, the mice may get additional boosts. Serumsamples may be periodically obtained from the mice by retro-orbitalbleeding for testing in ELISA assays to detect antibodies to thepolypeptide encoded by P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ IDNO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01(SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13),P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ IDNO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06(SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29),P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ IDNO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11(SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50),P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ IDNO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02(SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71),P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75).

[0312] After a suitable antibody titer has been detected, the animals“positive” for antibodies can be injected with a final intravenousinjection of the inmmunogen. Three to four days later, the mice aresacrificed and the spleen cells are harvested. The spleen cells are thenfused to a selected murine myeloma cell line such as P3X63AgU.1,available from ATCC, No. CRL 1597. The fusions generate hybridoma cellswhich can then be plated in 96 well tissue culture plates containing HAT(hypoxanthine, aminopterin, and thymidine) medium to inhibitproliferation of non-fused cells, myeloma hybrids, and spleen cellhybrids.

[0313] The hybridoma cells will be screened in an ELISA for reactivityagainst the protein encoded by P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01(SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13),P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ IDNO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06(SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29),P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ IDNO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11(SEQ ID NO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50),P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ IDNO:56), P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62) P00628_H02(SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71),P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75).

[0314] The positive hybridoma cells can be injected intraperitoneallyinto syngeneic Balb/c mice to produce ascites containing the antibodies.Antibodies are purified by ammonium sulfate precipitation, protein A orprotein G chromatography or other techniques well known in the art.

Example 8

[0315] Further Animal Models

[0316] The biological function of the P00184_D11 (SEQ ID NO:1),P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ IDNO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10(SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17),P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ IDNO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03(SEQ ID NO:29), P00223_F07 (SEQ ID NO:31), P00225_C01 (SEQ ID NO:32),P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ IDNO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_B04(SEQ ID NO:44), P00240_E05 (SEQ ID NO:45), P00241_E12 (SEQ ID NO:47),P00245_D06 (SEQ ID NO:48), P00246_D12 (SEQ ID NO:49), P00247_A04 (SEQ IDNO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10(SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00263_G06 (SEQ ID NO:60),P00267_F08 (SEQ ID NO:61), P00269_H08 (SEQ ID NO:62), P00312_C04 (SEQ IDNO:64), P00324_H02 (SEQ ID NO:65), P00628_H02 (SEQ ID NO:66), P00629_C08(SEQ ID NO:68), P00634_G11 (SEQ ID NO:70), P00641_G11 (SEQ ID NO:71),P00648_E12 (SEQ ID NO:73), and P00697_C03 (SEQ ID NO:75) genes and theencoded protein are further characterized in various animal models ofheart, kidney and inflammatory disorders.

1 84 1 1340 DNA Rattus norvegicus 1 gcggccgccc ctgacacaat ggctcagcttatgcctcagc gcagttcgct ccaccccaga 60 atggcatcct gcagaataca cggcccctcatccccatccc gcgccagaga caccggccag 120 cccactgtcc ccgccacaca ttaaacttgatcctcctaca cagacgcact cggagcagag 180 cgcttataca agcgcacagc cgtctccggcaccgccacac agacagatga tgccgccccg 240 accgacggcc agccccagac acaaccttctgaaaacacag aaaacaagtc ccagcccaag 300 cggctgcatg tgtccaacat ccccttccggttccgggatc cagacctccg acaaatgttt 360 ggccaatttg gtaaaatatt agatgttgaaattattttta atgagcgggg ctcgaaggga 420 tttggtttcg taactttcga aaatagtgcggatgcggaca gggcgaggga gaaattgcac 480 ggtaccgtgg tagagggccg taaaatcgaggttaataatg cgacagcacg cgtgatgact 540 aataaaaagg ccgtgaaccc ctacaccaatggctggaaat taaatccagt tgtgggcgcg 600 gtctacagcc ccgacttcta tgcaggcacggtgctgttgt gccaggccaa ccaggaggga 660 tcttccatgt acagtggccc cagttcacttgtatatactt ctgcaatgcc tggctttcca 720 tatccggccg ccactgctgc agctgcataccgaggggctc accttcgagg ccgtggtcgc 780 accgtgtaca acaccttcag agctgcggcgcccccacccc caatcccggc ctatggcgga 840 gtagtgtatc aagagccagt gtatggcaataaattgctac agggtggtta cgctgcatac 900 cgctacgccc agcccacccc tgccactgctgctgcctaca gtgacagtta cggacgagtt 960 tatgctgccg acccctacca ccacacacttgctccagccc ccacctacgg cgttggtgcc 1020 atgaatgctt ttgcgccctt gaccgatgccaagactagga gccatgctga tgatgtgggt 1080 ctcgttcttt cttcattgca ggctagtatataccaagggg gatacaaccg ttttgctcca 1140 tattaaatga taaaaccatt aaacaaacaagcaaaaaaca aaacaaaaac aaaaaaacca 1200 accttccaat gtggggagag aggaagctttccgaggcccg agtgttgcga cacatgcagt 1260 aggacatcac tttagcaact caaagaaacaacgaaaaaaa aaaaaaaaaa aaaaataagc 1320 ggccgaaggg gttcgctaga 1340 2 203PRT Rattus norvegicus 2 Met Thr Asn Lys Lys Ala Val Asn Pro Tyr Thr AsnGly Trp Lys Leu 1 5 10 15 Asn Pro Val Val Gly Ala Val Tyr Ser Pro AspPhe Tyr Ala Gly Thr 20 25 30 Val Leu Leu Cys Gln Ala Asn Gln Glu Gly SerSer Met Tyr Ser Gly 35 40 45 Pro Ser Ser Leu Val Tyr Thr Ser Ala Met ProGly Phe Pro Tyr Pro 50 55 60 Ala Ala Thr Ala Ala Ala Ala Tyr Arg Gly AlaHis Leu Arg Gly Arg 65 70 75 80 Gly Arg Thr Val Tyr Asn Thr Phe Arg AlaAla Ala Pro Pro Pro Pro 85 90 95 Ile Pro Ala Tyr Gly Gly Val Val Tyr GlnGlu Pro Val Tyr Gly Asn 100 105 110 Lys Leu Leu Gln Gly Gly Tyr Ala AlaTyr Arg Tyr Ala Gln Pro Thr 115 120 125 Pro Ala Thr Ala Ala Ala Tyr SerAsp Ser Tyr Gly Arg Val Tyr Ala 130 135 140 Ala Asp Pro Tyr His His ThrLeu Ala Pro Ala Pro Thr Tyr Gly Val 145 150 155 160 Gly Ala Met Asn AlaPhe Ala Pro Leu Thr Asp Ala Lys Thr Arg Ser 165 170 175 His Ala Asp AspVal Gly Leu Val Leu Ser Ser Leu Gln Ala Ser Ile 180 185 190 Tyr Gln GlyGly Tyr Asn Arg Phe Ala Pro Tyr 195 200 3 867 DNA Rattus norvegicusunsure (0)...(0) n = A, T, C, or G 3 tctagcgaac cccttcgcga aggggttcgcctgtgctggt gggcgcggtg gcccgaagcc 60 ttggactcac tgcaggactg tgcagggaaccactgtccaa gcatcgggct aatagggggc 120 gcctgcctcg gtttaccctt cagcgtctggtgaaatcccg cagcgtctag ggaaagatcc 180 gttctgctcc gcgagggaaa cagagccgttgaccatggtt gcaacgggca gtttgagcag 240 taagaacacg gccagcattt cagagttgctggacggtggc tctcaccctg ggagtctgct 300 aagtgatttc gactactggg attatgtcgtccctgagccc aacctcaacg aggtggtgtt 360 tgaagagaca acatgccaga atttggttaaaatgttggag aactgtctgt ccaagtcaaa 420 gcaaaccaaa ctcggttgct ctaaggtcctggttcctgag aaactgaccc agagaattgc 480 ccaagatgtc ctgcggctct catccacagagccctgcggc cttcggggct gtgttatgca 540 cgtgaacttg gaaattgaaa atgtgtgtaaaaagctggat aggattgtgt gtgatgctag 600 tgtggtgccg acctttgagc tcacgctggtgttcaagcag gagagctgct cctggaccag 660 cctcaaggac ttcttcttta gcggaggtcgcttctcgtcg ggccttaagc gaactctgat 720 cctcagctcg ggatttcgac ttgttaagaaaaaactgtac tctctgattg gaacgacagt 780 cattgaggag tgctgaggag gaaaaaacaattaaaggtcc ctaatgagtg gctaacaaaa 840 anaaaannnn nnnnnnnnnn ngcggnc 867 4193 PRT Rattus norvegicus 4 Met Val Ala Thr Gly Ser Leu Ser Ser Lys AsnThr Ala Ser Ile Ser 1 5 10 15 Glu Leu Leu Asp Gly Gly Ser His Pro GlySer Leu Leu Ser Asp Phe 20 25 30 Asp Tyr Trp Asp Tyr Val Val Pro Glu ProAsn Leu Asn Glu Val Val 35 40 45 Phe Glu Glu Thr Thr Cys Gln Asn Leu ValLys Met Leu Glu Asn Cys 50 55 60 Leu Ser Lys Ser Lys Gln Thr Lys Leu GlyCys Ser Lys Val Leu Val 65 70 75 80 Pro Glu Lys Leu Thr Gln Arg Ile AlaGln Asp Val Leu Arg Leu Ser 85 90 95 Ser Thr Glu Pro Cys Gly Leu Arg GlyCys Val Met His Val Asn Leu 100 105 110 Glu Ile Glu Asn Val Cys Lys LysLeu Asp Arg Ile Val Cys Asp Ala 115 120 125 Ser Val Val Pro Thr Phe GluLeu Thr Leu Val Phe Lys Gln Glu Ser 130 135 140 Cys Ser Trp Thr Ser LeuLys Asp Phe Phe Phe Ser Gly Gly Arg Phe 145 150 155 160 Ser Ser Gly LeuLys Arg Thr Leu Ile Leu Ser Ser Gly Phe Arg Leu 165 170 175 Val Lys LysLys Leu Tyr Ser Leu Ile Gly Thr Thr Val Ile Glu Glu 180 185 190 Cys 5874 DNA Rattus norvegicus 5 tctagcgaac cccttcggtg gacagaacag cctgagtcaggatgaaagct ctcagggctg 60 tcctcctgat cttgctactc agtggacagc cagggagcagctgggcacaa gaagctggcg 120 atgtggacct ggagctagag cgctacagct acgatgatgacggtgatgac gatgatgacg 180 atgatgaaga agaggaagag gaggagacca acatgatccctggcagcagg gacagagcac 240 cgcctctaca gtgctacttc tgccaagtgc ttcacagcggggagagctgc aacgagacac 300 agagatgctc cagcagcaag cccttctgta tcacagtcatctcccatggc aaaactgaca 360 caggtgtcct gacgacctac tccatgtggt gtactgatacctgccagccc atcgtgaaga 420 cagtggacag cacccaaatg acccagacct gttgccagtccacactctgc aatattccac 480 cctggcagag cccccaaatc cacaaccctc tgggtggccgggcagacagc cccttgaagg 540 gtgggaccag acatcctcaa ggtgacaggt ttagccacccccaggttgtc aaggttactc 600 atcctcagag tgatggggct cacttgtcta agggtggcaaggctaaccag ccccagggaa 660 atggggccgg attccctgca ggctggagca aatttggtaacgtagttctc ctgctcacct 720 tcctcaccag tctgtgggca tcaggggcct aaagactcgtcctcccccaa ccaggaccct 780 tcagcctttc ctccctgaca accagcttca gagaataaacttgaatgtct tttgccatct 840 aaaaaaaaaa aaaaaaaaaa aaaaagcggc cgcc 874 6236 PRT Rattus norvegicus 6 Met Lys Ala Leu Arg Ala Val Leu Leu Ile LeuLeu Leu Ser Gly Gln 1 5 10 15 Pro Gly Ser Ser Trp Ala Gln Glu Ala GlyAsp Val Asp Leu Glu Leu 20 25 30 Glu Arg Tyr Ser Tyr Asp Asp Asp Gly AspAsp Asp Asp Asp Asp Asp 35 40 45 Glu Glu Glu Glu Glu Glu Glu Thr Asn MetIle Pro Gly Ser Arg Asp 50 55 60 Arg Ala Pro Pro Leu Gln Cys Tyr Phe CysGln Val Leu His Ser Gly 65 70 75 80 Glu Ser Cys Asn Glu Thr Gln Arg CysSer Ser Ser Lys Pro Phe Cys 85 90 95 Ile Thr Val Ile Ser His Gly Lys ThrAsp Thr Gly Val Leu Thr Thr 100 105 110 Tyr Ser Met Trp Cys Thr Asp ThrCys Gln Pro Ile Val Lys Thr Val 115 120 125 Asp Ser Thr Gln Met Thr GlnThr Cys Cys Gln Ser Thr Leu Cys Asn 130 135 140 Ile Pro Pro Trp Gln SerPro Gln Ile His Asn Pro Leu Gly Gly Arg 145 150 155 160 Ala Asp Ser ProLeu Lys Gly Gly Thr Arg His Pro Gln Gly Asp Arg 165 170 175 Phe Ser HisPro Gln Val Val Lys Val Thr His Pro Gln Ser Asp Gly 180 185 190 Ala HisLeu Ser Lys Gly Gly Lys Ala Asn Gln Pro Gln Gly Asn Gly 195 200 205 AlaGly Phe Pro Ala Gly Trp Ser Lys Phe Gly Asn Val Val Leu Leu 210 215 220Leu Thr Phe Leu Thr Ser Leu Trp Ala Ser Gly Ala 225 230 235 7 817 DNARattus norvegicus unsure (0)...(0) n = A, T, C, or G 7 tctagcgaaccccttcgagc gaaccccttc ggccagtacc ctgagccctg gtccctcctg 60 gagctgccccacagctctga ctgtggactg agggatgtta ggcggatcac ctgagcctcc 120 agaggctcacactaatgagc gggcgctctc ttcttagcca ctgttgcatt tggttttcat 180 tgactcctgggcctcgtttg agtgacactg tccttgtctt ttgtttcaga gctctcccag 240 tgttagtggactcagatgag gaaattatga ccagatctga aatagctgaa aaaatgttct 300 cttcagaaaagataatgtga tcagggcccc agtgggtcca gtgtgcatgg gagcgcggtc 360 aggtgatgggaaaggcctgg ctctcgtcaa aactgacagc tgcgctatga tacatgtctc 420 actttgttgtcttggagatc tgtgtatgca ggtgaagaac tcaagtgtgg gagggtctgc 480 cgcctcagaaagccatcttt gaaacggact cataaagtca gttttgttgc cattaagttg 540 cctgattttggaaacaattt aagaagtgtt aaagacatgt gttcagatgc ctcttaggcg 600 gcagccacaggcatgccagg ttgtgtccct cagttttctc cagacaaaag aatctgcagc 660 tgggcgtggcggcacactac tggcagttga aagtctgtaa tttcaaggcc aagcctggtc 720 tacatagttccaggacaacc agagagatct acatagtgag accctgcctc aaaacacaga 780 aaccnnannanaaaaaaaaa aaaaaaaaag cggccgc 817 8 61 PRT Rattus norvegicus 8 Met SerGly Arg Ser Leu Leu Ser His Cys Cys Ile Trp Phe Ser Leu 1 5 10 15 ThrPro Gly Pro Arg Leu Ser Asp Thr Val Leu Val Phe Cys Phe Arg 20 25 30 AlaLeu Pro Val Leu Val Asp Ser Asp Glu Glu Ile Met Thr Arg Ser 35 40 45 GluIle Ala Glu Lys Met Phe Ser Ser Glu Lys Ile Met 50 55 60 9 755 DNARattus norvegicus unsure (0)...(0) n = A, T, C, or G 9 tctagcgaaccccttcgcac atgggttcct gctgaccaag gggacatggc tctgaagatg 60 atgaggctggttactcagca ggagtagctg agctgagctg gccctggagg ccctggaggc 120 cctggagtagggcccaggat gcaggtgcta atgtctatcc ccggcgctct tcttcccgac 180 tctaccatgggatgtaactc caggagcccc tgccatctcc cgtaccaaaa gactgtggct 240 tccgtgtctactcagaaatc agttctactt cgtaaacagt gtttaaaacc agactcattt 300 aatcagagtgaaggattgca gtccattggc ttcttagcac agaagcagct gataacacaa 360 gtaaaccccagcccttgaga ggtagaagca agaggatcag aggttcaagc gcatcctcgg 420 ctccatcacaagttcaaaag ccgcctgcac caaatgggag tccttgtctc aaaaaaaaaa 480 aaaaaaaaagcaaagaaagc aaaggactcg atgacatgat ttatagacaa aagcagtggg 540 agaaaatactaaagccccac tgagctgcca gccaggtgtc tgtgactaca ggtcttttat 600 ctgctcatatatatttttac aaaaaatgaa attcatattg gtcgctattt tgctggctgc 660 tttgctcccgatcaacatga tttgcacgtt ttttccatca ataaatgtgc catgatattt 720 ttaaaaaaaaaaaaaaaaaa aaaaaaaagg gcncc 755 10 79 PRT Rattus norvegicus 10 Met GlnVal Leu Met Ser Ile Pro Gly Ala Leu Leu Pro Asp Ser Thr 1 5 10 15 MetGly Cys Asn Ser Arg Ser Pro Cys His Leu Pro Tyr Gln Lys Thr 20 25 30 ValAla Ser Val Ser Thr Gln Lys Ser Val Leu Leu Arg Lys Gln Cys 35 40 45 LeuLys Pro Asp Ser Phe Asn Gln Ser Glu Gly Leu Gln Ser Ile Gly 50 55 60 PheLeu Ala Gln Lys Gln Leu Ile Thr Gln Val Asn Pro Ser Pro 65 70 75 11 806DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 11 tctagcgaaccccttcgcag ctctctgacc tgcgtcgccg ccgctctccg ctcttgattt 60 cgccgtgatgtcgaccgcaa tgaacttcgg gaccaaaagc ttccagccgc ggcccccaga 120 caaaggcagcttcccgctag accacttcgg tgagtgtaaa agctttaagg aaaaattcat 180 gaagtgtctccgcgacaaga actatgaaaa tgctctgtgc agaaatgaat ctaaagagta 240 tttaatgtgcaggatgcaaa ggcagctgat ggcaccagaa ccactagaga aactcggctt 300 tagagacataatggaggaga aaccggaggc aaaggacaaa tgttgagaat cactgggctg 360 tgtccccctacctggagcag agctgagccc ttctgcccac cgtggagaga gctgagccat 420 cctgtgctgcccagaggagg ggctctccgt gtcgactttg gctcatccct gcagcacaga 480 ccaaactgctttctctactg accacacttc tgcttcagag agnggtttct cctgtctgng 540 tgtggcacaggatctgctca nggctgaaca ctgatgtgat atgatatccc acctagtgtg 600 gccgcacaccaaaaggcctg gacaggattt cacagtgact caacctgagt cctcacaccc 660 ggaacctgtcagcgaaaacc aancgaagca aaatgnctgg cttttggctt acaaacccca 720 tnatttgntttcccttctct tgggtctttg ttttgacaaa nctggcatac aaagtnggaa 780 gggggaaataaaaaaaaaaa aaaaaa 806 12 92 PRT Rattus norvegicus 12 Met Ser Thr Ala MetAsn Phe Gly Thr Lys Ser Phe Gln Pro Arg Pro 1 5 10 15 Pro Asp Lys GlySer Phe Pro Leu Asp His Phe Gly Glu Cys Lys Ser 20 25 30 Phe Lys Glu LysPhe Met Lys Cys Leu Arg Asp Lys Asn Tyr Glu Asn 35 40 45 Ala Leu Cys ArgAsn Glu Ser Lys Glu Tyr Leu Met Cys Arg Met Gln 50 55 60 Arg Gln Leu MetAla Pro Glu Pro Leu Glu Lys Leu Gly Phe Arg Asp 65 70 75 80 Ile Met GluGlu Lys Pro Glu Ala Lys Asp Lys Cys 85 90 13 717 DNA Rattus norvegicusunsure (0)...(0) n = A, T, C, or G 13 tctagcgaac cccttcncga aggggttcgccgagaggtgg gagccaaaag gatggagcat 60 ccgccggtgg tggctggtgg ccgcaatcttggtggtcctg atcggggttg tcttagtctg 120 cctgatagtc tacttcgcca acgcagcgcacagcgaggcc tgtaagaacg ggttgcggtt 180 gcaggatgag tgccgaaaca ccacgcacctgttgaagcac cagctnaccc gcgcccagga 240 cagcctgctg cagacggaga tgcaggcaaactcctgcaac cagaccgtga tggaccttcg 300 ggattccctg aagaagaagg tgtctnaaacccaggagcaa cangcccgca tcaaggaact 360 tgagaataag atcgagaggc tgaaccaagagctggagaaa tttgaggacc caaaaggaaa 420 tttctaccac agtgcangtg aactcaagcgggttcgtggt ggncttcanc ctacttgtgc 480 tttgtggcgg gactgttctn cactttttangacccaataa ttgggangta caaacctgtg 540 taggcattgn nggtngtaat ggcttttgagggggtcctgg cacccttaag atgtgaanac 600 cattangnng gacccaaaat gnnttttcttgntttgaact ggggcggacc cggagtgggg 660 ggcnggaaat aanntattnn ggnnggaaanaaaaaaaaaa aaaaaaaaaa gcggccc 717 14 86 PRT Rattus norvegicus UNSURE(0)...(0) Xaa = any amino acid 14 Met Gln Ala Asn Ser Cys Asn Gln ThrVal Met Asp Leu Arg Asp Ser 1 5 10 15 Leu Lys Lys Lys Val Ser Xaa ThrGln Glu Gln Xaa Ala Arg Ile Lys 20 25 30 Glu Leu Glu Asn Lys Ile Glu ArgLeu Asn Gln Glu Leu Glu Lys Phe 35 40 45 Glu Asp Pro Lys Gly Asn Phe TyrHis Ser Ala Xaa Glu Leu Lys Arg 50 55 60 Val Arg Gly Gly Leu Xaa Pro ThrCys Ala Leu Trp Arg Asp Cys Ser 65 70 75 80 Xaa Leu Phe Xaa Thr Gln 8515 1235 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C or G 15tctagcgaac cccttcgccc agctgctaga agccaggctg gcctggtgag gcatgagcat 60gaagatgaac ccaggtgaca aggacaagat gttgctcttc tccccaccct ttgacccctg 120tcttctaagg catctaggaa ggaaccagtg tccttggtac tgatttactt agattcaacc 180taagggtcca gccactgact aaggccaagg ccatttttcc atacctggga gggtagagat 240tcagggttgt gggtaagtgg gcactaaaca tggatttgca agggaaaacg acagggcatc 300gagctaaatt tgaatttaca tgaaattctg aaatgtactt gtatgaagaa actgttatct 360gaaacctaac ttaaatgggc atcctgcctt ttgtctggtg agaaatgaaa gtgatctaca 420ataagtgtca aagcaacaag gcccctctgg atatgtctag gccaggatga ggatactaag 480tgccttcaaa gcgagaggga ggcaggccaa gaacactgcc ctactgaaag gcaggcttgg 540ccggctaggg cctccaaggc cctgatccct gaggcaccac agccacaact tgtgtaggcc 600tggcccaggt cagtgaatag gttctaggca gtggttctca accttcctaa tgctgcaacc 660cttcaataca gtttctcctg ttgtagtaat ccccaaccat aaaattattt tcattgcgac 720ttcataactg gacttttgct actgttatga atcataatgt aaatattttt tggagctaga 780ggtttaccaa gggggttgtg agccataggt tgaaaaccat tgttctagga atagctccag 840gggtggtttc tgaggccccc gcaaggtggg atctatgggg cagggttgga tcttctccaa 900gagcccccaa caggatatat atatatatat atatatatat atatatatat atatatatat 960atatactttg atagcatccc atggaacgac tgtctcctga tactaaaggg agcttggaag 1020aaaccaaggc tgagagaagt tgtagagtgg gaaggtaggc gaagggattg aggtgacaca 1080gtgatagccc cttcagggtg gggtctaccc nagacagcag ataaaggcct taggatggga 1140gattactctg gctgctcaga ggggaacaca gggacacagc accaataaaa tctctttctt 1200ttcaaaaaaa aaaaaaaaaa aaaaaaaagc ggncc 1235 16 36 PRT Rattus norvegicus16 Met Ser Met Lys Met Asn Pro Gly Asp Lys Asp Lys Met Leu Leu Phe 1 510 15 Ser Pro Pro Phe Asp Pro Cys Leu Leu Arg His Leu Gly Arg Asn Gln 2025 30 Cys Pro Trp Tyr 35 17 633 DNA Rattus norvegicus 17 tctagcgaaccccttcgatt ttattagctc ttgcttctcc attcctcata atttatgaat 60 tatacagccttcgcttgaat acgcgtctga agttatgctt tgtgttgttg tgggtttttt 120 tttttttttcttttcttttt ttttggagct ggggaccgaa cccagggcct tgttgctcta 180 ccactgagctaaatccccaa cccctgttgt gtgttttaaa taagtctctt actgtccatt 240 ttgtaattagtgttgttacc ttgtaataat agacatcata caaagtttcc tcttttttgt 300 gccagtgctgagaacatgag aaacatttaa tgagtatttg tttgttaaat aatatttata 360 acggctagaatggcagacac acatggtagc acatgatggt gattttcggg ggccttttgt 420 ttgctcagagctggtaatct ctgccggttg gtttgctttg cctggtctgg gactaacctc 480 acattttctcactcttgctt tccgagagat tagtcatcct tcctgtccta ctgggctctc 540 gatagcgctcatcagcatac tgcatttcaa tcccagcgaa ggggttcgcc gaaggggttc 600 gctaggccagtgtgatggat atctgcagaa ttc 633 18 83 PRT Rattus norvegicus 18 Met Val AlaHis Asp Gly Asp Phe Arg Gly Pro Phe Val Cys Ser Glu 1 5 10 15 Leu ValIle Ser Ala Gly Trp Phe Ala Leu Pro Gly Leu Gly Leu Thr 20 25 30 Ser HisPhe Leu Thr Leu Ala Phe Arg Glu Ile Ser His Pro Ser Cys 35 40 45 Pro ThrGly Leu Ser Ile Ala Leu Ile Ser Ile Leu His Phe Asn Pro 50 55 60 Ser GluGly Val Arg Arg Arg Gly Ser Leu Gly Gln Cys Asp Gly Tyr 65 70 75 80 LeuGln Asn 19 607 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G19 tctagcgaac cccttcgcct ttctccaaag ccttcccgtt tcctcttgac agctacgggc 60tgaggcagcc attcctgcag cagcgctcgg ccggtgaagg gccgaactga cgcctcctag 120atctgtctcg gctgaattac tctcacccgt ttccattctg tgtgcaccag aaatctgaga 180tccaggagta tcaacagcaa agatgtctaa tgagccaccc cctccttatc caggagggcc 240tacagcccca ctactggagg aaaaaagtgg agccccacat accccaggcc gaacctttcc 300agctgtgatg cagccaccac caggcatgcc actgccctct gttgacattg cccccccgcc 360ctatgagccg cctggccatc cagggcctaa gcctggtttw atgcccccca cnttaccaca 420cattcnaana accttnntnt gtaaaagtta aataanaang gagggattcg anccccctnc 480aacnggtttc aagccaattt ymtaaccatt ttgttttttt cwtttaaaaa aaaaaaaaaa 540aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa ggggaaaaaa aaaaaaaaaa aaaaaaaggg 600gggcccc 607 20 82 PRT Rattus norvegicus UNSURE (0)...(0) Xaa = any aminoacid 20 Met Ser Asn Glu Pro Pro Pro Pro Tyr Pro Gly Gly Pro Thr Ala Pro1 5 10 15 Leu Leu Glu Glu Lys Ser Gly Ala Pro His Thr Pro Gly Arg ThrPhe 20 25 30 Pro Ala Val Met Gln Pro Pro Pro Gly Met Pro Leu Pro Ser ValAsp 35 40 45 Ile Ala Pro Pro Pro Tyr Glu Pro Pro Gly His Pro Gly Pro LysPro 50 55 60 Gly Xaa Met Pro Pro Thr Leu Pro His Ile Xaa Xaa Thr Xaa XaaCys 65 70 75 80 Lys Ser 21 1456 DNA Rattus norvegicus unsure (0)...(0) n= A, T, C, or G 21 tctagcgaac cccttcgcaa agtcctaagc cttacatgagaaaatttaag acacccttaa 60 tgattgcgga agaaaaatac agacaacaaa gggaagagcttgagaaacag agacgggaga 120 gttcttgcca tagcatcatc aaaacagaaa cccagcaccgcagcttatca gagaaagaga 180 aagaaacaga gttacaaaaa gcagctgagg caatgtccactcccagaaag gattcagact 240 tcactagggc acagcccaac ctggaaccta aaagcaaggctgtgatcgcc agtgaatgct 300 ctgaaagcca gctctctaca gcttccgcat tgacagtcgctaccgagagg ctccagcatg 360 ttctagccgc ttcagacgat aagcttaccc tgcgacgggaaggcacacag aactcaagtg 420 acaccctaca atcgaaaaca gcttgtgaga ttaaccagagtcacaaggaa tgtaggacag 480 agcaaacatt tgagcaacac gtggagaagt tgcccttcccccaaaccaaa cccatttccc 540 cgagtttcaa agtgaaaact atcaggcttc cagctctagatcatacgctg actgaaacag 600 atctcagttc tgaacgccgc gtaaagcaat ccgaaattgacgttcaaacc agtactaaag 660 aaatgaataa ggaaattaag aaaaccgaag tgagcacacagtgtgataat aagcaatctg 720 tggctgaaaa atattttcaa ttacctaaaa cagagaaacgggtgacggta caaatgccca 780 aagactatgc agcgaaaagt catcaaagca aactccaaacagttcccaag aagcatggag 840 gattggggga gtttgacaga gggaatgtcc tggggagggaaggaaaaaat caggactcct 900 ccatgagcag tacaaaagaa agcagggtaa tagttgaaagaaagcaagaa catctacagg 960 accagagcgt accaaggtta gtccaacaaa agattatcggtgaaagcctg gactcacggg 1020 ttcagaattt tcagcagaca caaacacaaa cttctaggattgagcataaa gaactgtccc 1080 aaccttacag tgagaaaaaa tgtcttagag acaaggacaaacaacaaaaa caggtctcct 1140 ctaacactga cgattcaaag caagagataa cacaaaaacaatcttcattt tcctctgtga 1200 gagaatccca gcaggatgga gaaaaatgtg ccataaaaatattggaattc ttgagaaaac 1260 gtgaagaact acagcagatt ttgtctaggg taaaacagtttgaagcagat tcaaataaaa 1320 gtggccttaa aacatttcag acactgttaa atattgctccggtgtggctg ataagtgagg 1380 agaaaagaga atatggagtt cgtgttgcca tggagaataattagaaaaaa taaaaaaaaa 1440 aaaaaaaagc ggcgnc 1456 22 462 PRT Rattusnorvegicus 22 Met Arg Lys Phe Lys Thr Pro Leu Met Ile Ala Glu Glu LysTyr Arg 1 5 10 15 Gln Gln Arg Glu Glu Leu Glu Lys Gln Arg Arg Glu SerSer Cys His 20 25 30 Ser Ile Ile Lys Thr Glu Thr Gln His Arg Ser Leu SerGlu Lys Glu 35 40 45 Lys Glu Thr Glu Leu Gln Lys Ala Ala Glu Ala Met SerThr Pro Arg 50 55 60 Lys Asp Ser Asp Phe Thr Arg Ala Gln Pro Asn Leu GluPro Lys Ser 65 70 75 80 Lys Ala Val Ile Ala Ser Glu Cys Ser Glu Ser GlnLeu Ser Thr Ala 85 90 95 Ser Ala Leu Thr Val Ala Thr Glu Arg Leu Gln HisVal Leu Ala Ala 100 105 110 Ser Asp Asp Lys Leu Thr Leu Arg Arg Glu GlyThr Gln Asn Ser Ser 115 120 125 Asp Thr Leu Gln Ser Lys Thr Ala Cys GluIle Asn Gln Ser His Lys 130 135 140 Glu Cys Arg Thr Glu Gln Thr Phe GluGln His Val Glu Lys Leu Pro 145 150 155 160 Phe Pro Gln Thr Lys Pro IleSer Pro Ser Phe Lys Val Lys Thr Ile 165 170 175 Arg Leu Pro Ala Leu AspHis Thr Leu Thr Glu Thr Asp Leu Ser Ser 180 185 190 Glu Arg Arg Val LysGln Ser Glu Ile Asp Val Gln Thr Ser Thr Lys 195 200 205 Glu Met Asn LysGlu Ile Lys Lys Thr Glu Val Ser Thr Gln Cys Asp 210 215 220 Asn Lys GlnSer Val Ala Glu Lys Tyr Phe Gln Leu Pro Lys Thr Glu 225 230 235 240 LysArg Val Thr Val Gln Met Pro Lys Asp Tyr Ala Ala Lys Ser His 245 250 255Gln Ser Lys Leu Gln Thr Val Pro Lys Lys His Gly Gly Leu Gly Glu 260 265270 Phe Asp Arg Gly Asn Val Leu Gly Arg Glu Gly Lys Asn Gln Asp Ser 275280 285 Ser Met Ser Ser Thr Lys Glu Ser Arg Val Ile Val Glu Arg Lys Gln290 295 300 Glu His Leu Gln Asp Gln Ser Val Pro Arg Leu Val Gln Gln LysIle 305 310 315 320 Ile Gly Glu Ser Leu Asp Ser Arg Val Gln Asn Phe GlnGln Thr Gln 325 330 335 Thr Gln Thr Ser Arg Ile Glu His Lys Glu Leu SerGln Pro Tyr Ser 340 345 350 Glu Lys Lys Cys Leu Arg Asp Lys Asp Lys GlnGln Lys Gln Val Ser 355 360 365 Ser Asn Thr Asp Asp Ser Lys Gln Glu IleThr Gln Lys Gln Ser Ser 370 375 380 Phe Ser Ser Val Arg Glu Ser Gln GlnAsp Gly Glu Lys Cys Ala Ile 385 390 395 400 Lys Ile Leu Glu Phe Leu ArgLys Arg Glu Glu Leu Gln Gln Ile Leu 405 410 415 Ser Arg Val Lys Gln PheGlu Ala Asp Ser Asn Lys Ser Gly Leu Lys 420 425 430 Thr Phe Gln Thr LeuLeu Asn Ile Ala Pro Val Trp Leu Ile Ser Glu 435 440 445 Glu Lys Arg GluTyr Gly Val Arg Val Ala Met Glu Asn Asn 450 455 460 23 2023 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 23 gaattgtaat acgactcactatagggcgaa ttgggcccct agcgaacccc ttcgacaaca 60 tcaaagagga cagatctaaccctagactga ggccggaggc ctggaccaat tacctgaggg 120 atgtccacag agcctttgcactgctgaaca gtcaccctga tccaaaccaa gtaaatggga 180 ctccaactgc accaagcagtggcctcccag tcacctctgc tgagctcttg gtgccggcag 240 agatggcttc tgcagagtcaggtgaagacc caagtcatgt ggttggggaa acgcctcctt 300 tgaccttgcc agccaacctccaaaccctgc atccgaacag accaacgttg agtccagaga 360 gaaaacttga atggaataacgacattccag aagtgaatcg tttgaattct gaacactgga 420 gaaaaactga ggagcagccaggacgggggg aggtgcttct ccccgaaggt gacgtcagtg 480 gcaacggtat gacagagctgttgcccatcg gtcggcacca acaaaagcgt ccccacgatg 540 cggggccaga ggaccatgcttttgaagatc aattgcatcc tctcgtccac tctgacagaa 600 ctcccgttca tcgggtgttcgatgtgtccc acttggagca gcctgttcac tccagccacg 660 tggaaggaat gttggccaagatggagggga tggcacaaag gagtgggcac caagtctcga 720 aggcagcgcc tcctctccagtcacttcttg cttagattac atgttgccta acaatgtttc 780 tttccatgtt ttgattagtaaactaactcg tggtggcaat catgactccc aaccttctga 840 gctcccccgg gtacgcttgcaccgtagacg ctcatgtgcg caccgtgcgg gtgatgctca 900 cacacagact cattgtaattcaccgtttta ccgagaaggg ggggggggcg aattttctgt 960 gttgatgctt tgtttttggtactaaaacag nattatcttt tgaatattgt agggacatga 1020 gtatataaag tctatccagtcaaaatggct agaattgngc ctttgtaagt tttaaaaact 1080 tgatgcccac atgagtctgtgagcacatnt ttcccgcctg cctaacggag ttggaatttg 1140 tttctaacca ctgtaattcttcaacatcat cacctttggt tcagtgattt tgcactttga 1200 gtttggatac tgtgtctgcttggttggtag tgttagtatt tttcttttaa acaggcttat 1260 cagagttgca cactttgtcctaggcagggc aaaggaatag acgcccagca aggacacaca 1320 gtataggtaa catactgcttatcgtacgct tttcccacaa agcattgcat gtgtttttac 1380 ctcgacgtgc taaagttgattagcagaaag gcatgactca caattttggt ggtaaaaaat 1440 aaaccctgag ggagcaagcaataactaaaa caagattgag ctgctctctc tgtgcttact 1500 aaatagatgc tcgccctgctaatgcttgcc ctcttgaaag aagaaacagg atgcacactg 1560 ctttatttca atcttcctctttttttcttg gtttcaccag tgagcgtaag cattggaaaa 1620 atatgtgtag tcttatctttctataagacg attttaataa actaaaatca caaatgctgt 1680 aaagtttgtg cgcaccagaatggaggctaa cttcataaac attgtgctgt gcgaatattc 1740 ctaaaatgat ccccaagctgtggttttcta gaagacatag ttcagaaccg cttttgaaaa 1800 atctgtcctc gtgagctcactcagtttctg tcggactttt agagacagtg gaaggattac 1860 ctcattgaga cgtttccgtgtcctcttcaa ctccacaggg tcttgacggt ggctttgttt 1920 ttccttctag actattcaaacatgtagata agttatattt ttctttaagt gtttaaagta 1980 aacacttttc aaaaaaaaaaaaaaaaaaaa aaaaagcggc cgc 2023 24 170 PRT Rattus norvegicus 24 Met AlaSer Ala Glu Ser Gly Glu Asp Pro Ser His Val Val Gly Glu 1 5 10 15 ThrPro Pro Leu Thr Leu Pro Ala Asn Leu Gln Thr Leu His Pro Asn 20 25 30 ArgPro Thr Leu Ser Pro Glu Arg Lys Leu Glu Trp Asn Asn Asp Ile 35 40 45 ProGlu Val Asn Arg Leu Asn Ser Glu His Trp Arg Lys Thr Glu Glu 50 55 60 GlnPro Gly Arg Gly Glu Val Leu Leu Pro Glu Gly Asp Val Ser Gly 65 70 75 80Asn Gly Met Thr Glu Leu Leu Pro Ile Gly Arg His Gln Gln Lys Arg 85 90 95Pro His Asp Ala Gly Pro Glu Asp His Ala Phe Glu Asp Gln Leu His 100 105110 Pro Leu Val His Ser Asp Arg Thr Pro Val His Arg Val Phe Asp Val 115120 125 Ser His Leu Glu Gln Pro Val His Ser Ser His Val Glu Gly Met Leu130 135 140 Ala Lys Met Glu Gly Met Ala Gln Arg Ser Gly His Gln Val SerLys 145 150 155 160 Ala Ala Pro Pro Leu Gln Ser Leu Leu Ala 165 170 251802 DNA Rattus norvegicus 25 tctagcgaac cccttcgggg gttttcatcatggagctgtc gcggcggatt tgtctcgtcc 60 gactgtggct gttgctactg tcattcttactgggcttcag cgcgggatct gccctcaact 120 ggcgggaaca agaaggcaag gaagtatgggattacgtgac tgttcgagag gatgcacgca 180 tgttctggtg gctctactat gccaccaacccttgcaagaa cttctcagag ctgcctctgg 240 tcatgtggct tcagggtggt ccaggtggttctagcactgg atttggaaac tttgaggaaa 300 tcggccctct tgacacccga ctcaagccacggaacactac ctggctgcag tgggccagtc 360 tcctgttcgt ggacaatcct gtgggcacgggcttcagtta cgtgaacacg acagatgcct 420 acgcaaagga cctggacacg gtggcttccgacatgatggt cctcctgaaa tccttctttg 480 attgtcataa agaattccag acggttccgttctacatttt ctcagaatcc tacggaggaa 540 agatggctgc tggcatcagt ttagaacttcacaaggctat tcagcaaggg accatcaagt 600 gcaacttctc tggggttgct ttgggtgactcctggatctc ccctgtggat tcagtgctgt 660 cctggggacc ttacctgtac agcgtgtctctccttgataa taaaggcttg gctgaggtgt 720 ccgacattgc ggagcaagtc ctcaatgaaaaacaagggct tctacaagga agccactcag 780 ctgtggggga aagcagaaat gatcattgaaaagaacaccg acggggtaaa cttctataac 840 atcttaacta aaagcacccc cgacacctctatggagtcga gcctcgagtt cttccggagc 900 cccttagttc gtctctgtca gcgccacgtgagacacctac aaggagacgc cttaagtcag 960 ctcatgaacg gtcccatcaa aaagaagctcaaaattatcc ctgacgacgt ctcctgggga 1020 gcccagtcgt cctccgtctt cataagcatggaagaggact tcatgaagcc tgtcatcgac 1080 atcgtggata cgttgctgga actcggggtcaatgtgactg tgtacaatgg gcagctggat 1140 ctcattgtgg acaccatagg tcaggagtcctgggttcaga agctgaagtg gccacagctg 1200 tccagattca atcagctaaa atggaaggccctgtacaccg atcctaagtc ttcagaaaca 1260 tctgcatttg tcaagtccta tgagaacctagcgttctact ggatcctaaa ggcgggtcac 1320 atggttcctg ctgaccaagg ggacatggctctgaagatga tgaggctggt tactcagcag 1380 gagtagctga gctgagctgg ccctggaggccctggaggcc ctggagtagg gcccaggatg 1440 caggtgctaa tgtctatccc cggcgctcttcttcccgact ctaccatggg atgtaactcc 1500 aggagcccct gccatctccc gtaccaaaagactgtggctt ccgtgtctac tcagaaatca 1560 gttctacttc gtaaacagtg tttaaaaccagactcattta atcagagtga aggattgcag 1620 tccattggct tcttagcaca gaagcagctgataacacaag taaaccccag cccttgagag 1680 gtagaagcaa gaggatcaga ggttcaagcgcatcctcggc tccatcacaa gttcaaaagc 1740 cgcctgcacc aaatgggagt ccttgtctcaaaaaaaaaaa aaaaaaaaaa aaaagcggcc 1800 gc 1802 26 259 PRT Rattusnorvegicus 26 Met Glu Leu Ser Arg Arg Ile Cys Leu Val Arg Leu Trp LeuLeu Leu 1 5 10 15 Leu Ser Phe Leu Leu Gly Phe Ser Ala Gly Ser Ala LeuAsn Trp Arg 20 25 30 Glu Gln Glu Gly Lys Glu Val Trp Asp Tyr Val Thr ValArg Glu Asp 35 40 45 Ala Arg Met Phe Trp Trp Leu Tyr Tyr Ala Thr Asn ProCys Lys Asn 50 55 60 Phe Ser Glu Leu Pro Leu Val Met Trp Leu Gln Gly GlyPro Gly Gly 65 70 75 80 Ser Ser Thr Gly Phe Gly Asn Phe Glu Glu Ile GlyPro Leu Asp Thr 85 90 95 Arg Leu Lys Pro Arg Asn Thr Thr Trp Leu Gln TrpAla Ser Leu Leu 100 105 110 Phe Val Asp Asn Pro Val Gly Thr Gly Phe SerTyr Val Asn Thr Thr 115 120 125 Asp Ala Tyr Ala Lys Asp Leu Asp Thr ValAla Ser Asp Met Met Val 130 135 140 Leu Leu Lys Ser Phe Phe Asp Cys HisLys Glu Phe Gln Thr Val Pro 145 150 155 160 Phe Tyr Ile Phe Ser Glu SerTyr Gly Gly Lys Met Ala Ala Gly Ile 165 170 175 Ser Leu Glu Leu His LysAla Ile Gln Gln Gly Thr Ile Lys Cys Asn 180 185 190 Phe Ser Gly Val AlaLeu Gly Asp Ser Trp Ile Ser Pro Val Asp Ser 195 200 205 Val Leu Ser TrpGly Pro Tyr Leu Tyr Ser Val Ser Leu Leu Asp Asn 210 215 220 Lys Gly LeuAla Glu Val Ser Asp Ile Ala Glu Gln Val Leu Asn Glu 225 230 235 240 LysGln Gly Leu Leu Gln Gly Ser His Ser Ala Val Gly Glu Ser Arg 245 250 255Asn Asp His 27 630 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C,or G 27 tctagcgaac cccttcgcga aggggttcgc taggttgcgt ttgtggagaaaaatctgttc 60 tacctcaggg ctgtgagaac ggcactcctg atgtctgaga aagagaaacaagattggctg 120 aaggatcctc cgttccttca gagacctggg tggagagcat tagggacacgaagaacagag 180 tagcggaaga agagttctta agtaataagt ttacctcctg actggctcacatcactgcct 240 tactctgtag aaagcaggtc atctcatgga tttccccctc ccacccccccagctggatca 300 ttttttgact cagggaaaat aattaaatta ttgtccaact gttagtgttgatcggtaaca 360 gcagaaaggc agaaagttcc tgataatctc aatattatct tttcaaaagtattttcctgg 420 aatgttgttt gctttggcat tacaaagttc tgtactctta aaaatattttgacttgctgg 480 gcatggaggt cacaccttta atccagaggc aggcatggat ccacaggagttcaaggccgc 540 ctggctacaa agcgagttca agggcagcca gggctacaca gagagaccttgtctcntnac 600 cnntnannaa aaaacnaaaa agccggccgc 630 28 30 PRT Rattusnorvegicus 28 Met Ser Glu Lys Glu Lys Gln Asp Trp Leu Lys Asp Pro ProPhe Leu 1 5 10 15 Gln Arg Pro Gly Trp Arg Ala Leu Gly Thr Arg Arg ThrGlu 20 25 30 29 445 DNA Rattus norvegicus 29 tctagcgaac cccttcggtatagtctttag gtagtggctt agtccctgga agctctggtt 60 gcttggcatt tcaacgtgcttcttaaataa ctgttttatt agtcagtaca agatgctttg 120 tatatcagat ctgaaatatcttaaaattat cacttgcatt gtaaattact attcctttcg 180 cagaaataat gaatgcttcaagaaaaaaaa aagctgtttg tattgggttt aaaacgtttc 240 caaacaccaa ttattctttacttaagtcat ccgatctagt tattaaatta ttattactgc 300 cttcacacta tcaaagatggtaaatatctg atagaatcat attcaaaata cttctgtttc 360 acatttcttg agaaagtactgactgtctga gttctttctc aagaaatgtg aaacagaagt 420 attttgaatc gaaggggttcgctag 445 30 39 PRT Rattus norvegicus 30 Met Leu Cys Ile Ser Asp Leu LysTyr Leu Lys Ile Ile Thr Cys Ile 1 5 10 15 Val Asn Tyr Tyr Ser Phe ArgArg Asn Asn Glu Cys Phe Lys Lys Lys 20 25 30 Lys Ser Cys Leu Tyr Trp Val35 31 273 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 31tctagcgaac cccttcggaa gaactgtata tttgtgcctt gttctgcaag ttaaaaagct 60ggtccagaca gtgtcataga attaactttt catttctgta ttaattttag gactgcaaaa 120atcccaaagc tgtatactta gattggattc aataaaaagt ttaagtttac tnaanaaaaa 180aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaanaaaaa aaaaaaaagg 240aaaaaaaaaa ncggncnnaa aaaaggnggc cgc 273 32 2077 DNA Rattus norvegicus32 tctagcgaac cccttcgggg gaacccaagc ggcttcgccc aggcattcgc gcgggcgccc 60gcggtctggg tcccacctcc tctgctttcg cacccttgaa gttttggagc accaggaaaa 120gagggcaagg aaggagaggg gaagcgaaag catatcctaa aacatttact taaaggagga 180aagaaaaggg gtcgcagaaa tggctggggc aattatagaa aacatgagca ccaagaagct 240ctgcattgtt ggagggattc ttctggtttt ccaaatcgtt gcctttctgg tgggaggctt 300gatcgctcca gcacccacaa cggcagtgtc ctacgtggca gcaaaatgtg tggatgtccg 360gaagaaccac cataaaacaa gatggctgat gccctgggga ccaaacaagt gtaacaagat 420caatgacttc gaagaagcaa ttccaaggga aattgaagcg aatgacattg tgttttctgt 480acacattccc ctcccttcta tggagatgag cccatggttc cagtttatgc tgtttatcct 540gcagatagac attgctttca agctaaacaa ccaaatcaga gaaaatgcag aagtttccat 600ggatgtttcc ctgggttacc gtgatgatat gttttctgag tggactgaaa tggcgcacga 660aagagtacca cgtaaactca gatgcacttt cacatccccc aagaccccag agcatgaagg 720tcgtcattat gaatgtgatg tccttccttt catggaaatt gggtcagtgg ctcataagta 780ttaccttcta aatatccggc tacctgtaaa tgagaagaag aaaatcaatg ttggaattgg 840ggaaataaag gacattcggt tggtgggaat ccaccaaaat ggaggtttca ctaaggtatg 900gtttgctatg aagaccttcc tcacacccag catcttcatc attatggtgt ggtattggag 960aaggatcacc atgatgtccc gacctccagt gcttctggaa aaagtcatct ttgcccttgg 1020gatttccatg acctttatca atatccctgt ggaatggttt tccattggat ttgattggac 1080ctggatgctg ttatttggtg acatacgaca gggcatcttc tatgcaatgc ttctttcctt 1140ctggatcatc ttctgtggcg agcacatgat ggatcaacat gagcggaatc acattgcagg 1200gtattggaag caagttggac caattgctgt tggctctttc tgcctcttca tatttgacat 1260gtgtgagaga ggagtgcaac tcacaaatcc tttctacagt atctggacta cagatgttgg 1320aacagaactg gctatggctt tcatcattgt ggcaggtatc tgcctctgcc tctacttcct 1380gtttctgtgt ttcatggtat ttcaagtatt cagaaacatc agtgggaaac agtctagcct 1440cccagccatg agcaaagtcc ggaggctgca ctatgagggt ctgattttca ggttcaagtt 1500cctcatgctg atcaccttgg cttgtgctgc catgactgtt atcttcttca ttgttagtca 1560ggtgacagaa ggccattgga aatggggtgg ggtcacagtt caagtgagca gtgctttctt 1620cactggaatc tatgggatgt ggaacctgta tgtctttgct ttgatgttct tgtatgcacc 1680atcccataag aactatgggg aagaccagtc taatggtgac ctgggtgtcc acagcgggga 1740agaactgcag ctcactacca caatcaccca tgtagatgga ccgactgaga tctacaagtt 1800gacccgtaaa gaagcacagg agtagtaggc tatggcattc atcctcaggg caggtgatga 1860agccaagttg ctggtgcatg ctgaccctca tgaatatgct ttcgtatctt tatgtcccag 1920gatcattttt atcctgtcac gtttacaaga acatttctga catgcatacg tttactttta 1980ccatgtatta gttactttta tatttctgtg ataaaacacc atgagaaata caatttacag 2040aagcaaaaaa aaaaaaaaaa aaaaaaaaag cggccgc 2077 33 541 PRT Rattusnorvegicus 33 Met Ala Gly Ala Ile Ile Glu Asn Met Ser Thr Lys Lys LeuCys Ile 1 5 10 15 Val Gly Gly Ile Leu Leu Val Phe Gln Ile Val Ala PheLeu Val Gly 20 25 30 Gly Leu Ile Ala Pro Ala Pro Thr Thr Ala Val Ser TyrVal Ala Ala 35 40 45 Lys Cys Val Asp Val Arg Lys Asn His His Lys Thr ArgTrp Leu Met 50 55 60 Pro Trp Gly Pro Asn Lys Cys Asn Lys Ile Asn Asp PheGlu Glu Ala 65 70 75 80 Ile Pro Arg Glu Ile Glu Ala Asn Asp Ile Val PheSer Val His Ile 85 90 95 Pro Leu Pro Ser Met Glu Met Ser Pro Trp Phe GlnPhe Met Leu Phe 100 105 110 Ile Leu Gln Ile Asp Ile Ala Phe Lys Leu AsnAsn Gln Ile Arg Glu 115 120 125 Asn Ala Glu Val Ser Met Asp Val Ser LeuGly Tyr Arg Asp Asp Met 130 135 140 Phe Ser Glu Trp Thr Glu Met Ala HisGlu Arg Val Pro Arg Lys Leu 145 150 155 160 Arg Cys Thr Phe Thr Ser ProLys Thr Pro Glu His Glu Gly Arg His 165 170 175 Tyr Glu Cys Asp Val LeuPro Phe Met Glu Ile Gly Ser Val Ala His 180 185 190 Lys Tyr Tyr Leu LeuAsn Ile Arg Leu Pro Val Asn Glu Lys Lys Lys 195 200 205 Ile Asn Val GlyIle Gly Glu Ile Lys Asp Ile Arg Leu Val Gly Ile 210 215 220 His Gln AsnGly Gly Phe Thr Lys Val Trp Phe Ala Met Lys Thr Phe 225 230 235 240 LeuThr Pro Ser Ile Phe Ile Ile Met Val Trp Tyr Trp Arg Arg Ile 245 250 255Thr Met Met Ser Arg Pro Pro Val Leu Leu Glu Lys Val Ile Phe Ala 260 265270 Leu Gly Ile Ser Met Thr Phe Ile Asn Ile Pro Val Glu Trp Phe Ser 275280 285 Ile Gly Phe Asp Trp Thr Trp Met Leu Leu Phe Gly Asp Ile Arg Gln290 295 300 Gly Ile Phe Tyr Ala Met Leu Leu Ser Phe Trp Ile Ile Phe CysGly 305 310 315 320 Glu His Met Met Asp Gln His Glu Arg Asn His Ile AlaGly Tyr Trp 325 330 335 Lys Gln Val Gly Pro Ile Ala Val Gly Ser Phe CysLeu Phe Ile Phe 340 345 350 Asp Met Cys Glu Arg Gly Val Gln Leu Thr AsnPro Phe Tyr Ser Ile 355 360 365 Trp Thr Thr Asp Val Gly Thr Glu Leu AlaMet Ala Phe Ile Ile Val 370 375 380 Ala Gly Ile Cys Leu Cys Leu Tyr PheLeu Phe Leu Cys Phe Met Val 385 390 395 400 Phe Gln Val Phe Arg Asn IleSer Gly Lys Gln Ser Ser Leu Pro Ala 405 410 415 Met Ser Lys Val Arg ArgLeu His Tyr Glu Gly Leu Ile Phe Arg Phe 420 425 430 Lys Phe Leu Met LeuIle Thr Leu Ala Cys Ala Ala Met Thr Val Ile 435 440 445 Phe Phe Ile ValSer Gln Val Thr Glu Gly His Trp Lys Trp Gly Gly 450 455 460 Val Thr ValGln Val Ser Ser Ala Phe Phe Thr Gly Ile Tyr Gly Met 465 470 475 480 TrpAsn Leu Tyr Val Phe Ala Leu Met Phe Leu Tyr Ala Pro Ser His 485 490 495Lys Asn Tyr Gly Glu Asp Gln Ser Asn Gly Asp Leu Gly Val His Ser 500 505510 Gly Glu Glu Leu Gln Leu Thr Thr Thr Ile Thr His Val Asp Gly Pro 515520 525 Thr Glu Ile Tyr Lys Leu Thr Arg Lys Glu Ala Gln Glu 530 535 54034 755 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 34tctaacgaac cccttcggag cgatggaatg agaaaggccc agaatgtgtt aagtctgtgc 60aggggaagtg tcctgagggg agggtctttg ggagggtcga aggccaggat ggcaaagtga 120aggtagctga ggttgcagtc ttgggtgccc actgctgtgc atctgtctgg ttatctaccc 180ctactttggg ctgacaactg cagggttggg tgtaggctgt ctcactgcat gccgggaagc 240tggagaagct ccacgggaac attgagggcc atggctttga gacactgcag agcatccttg 300gtctctgtaa ccacgtcacc taaccctgac aattccagac ccttcttcca ttgtccttgt 360gaaccatttg ggcttatctt tccctcttag tcgcaagggt caaaccaagg gtcagtcaag 420tagatgactg tcaccttggg cctccccaga ctctgctgcc ggggttggga gaccaaagta 480gaaactgcca ctacaaggcc ccaggatgag gtctctgttc tgtggacctg ctccccagat 540acaggcctca gacccatagg acgtggccgg tgctcaggga cacccaatcc ccggcctcac 600tccatcgagt actgacttct ttctctagtg ccttgggggt ctccatcctt cagttatggt 660atgaagaatc tatgcaaact gtataagctt ctgctcacca ataaacgctt tatttaaagc 720ttannnnnnn nnnannnnnn nnnnnaagcg gncgc 755 35 30 PRT Rattus norvegicus35 Met Arg Lys Ala Gln Asn Val Leu Ser Leu Cys Arg Gly Ser Val Leu 1 510 15 Arg Gly Gly Ser Leu Gly Gly Ser Lys Ala Arg Met Ala Lys 20 25 3036 1310 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 36tctagcgaac cccttcgcag aaacccaaag ttacagacca gaccctaccc aacatccagt 60cagcaatcca gctggagaaa cgcttgagat gacaagggac tttcagaagc aagccttgat 120aagacaggaa aagcagaatt ctaataaaga tatgaggaaa aatgacatgg gccttcaacc 180tctgcctgta gggaaggacg cacacagtgc accaggagtg acagtctctg ggaaaaacca 240caaaagaact caggcacctg acaagaaaca gagaattgat gtttgtctag aaagccagga 300ctttctaatg aagacaaata cttccaagga gttaaaaatg gcaatggaga ggtcctttaa 360tccagtcaac ctttccctga ctgtggtgta aaagaaaatg aggacgccct tctctccatc 420ttcccctcct tcttctcctt ccaattgcgt catctgaaat tgaatttcct ctcctcctcc 480accacctata atgctgtgcc tgaaaaaaat gagtttcctc cctcatcacc cacagagaag 540tcaagggctg aacttgagag cctcccaacc ctgcctcttc ctccaccacc aggagatgag 600aaatctgatc aggaatgtct accaacatcc ctacctcctc cccctcccac agctccatcc 660caaccagcac atcttctttc ctcctctgtt ctagaacatc acagtgaagc atttttacaa 720cagtattccc gaaaagaaac cttggactct catcggcttc actcacaggc taaaatccta 780acaggaaaat caccaccccc aacactcccc aaacccaaac ttcccgagag aatcaaagct 840aagatgagcc aggattcacc aagcggtgaa ttggaaagat ctctgtcaga tgtggaaatt 900aaaactaccc tctcaaagga tcagaaaagt tcgctggtgg cagaaagccg tgagcacaca 960gaggccaagc aagaagtatt ccgaaaaagc cttggaagaa aacagctgtc cattagctct 1020gcaaactccc tctctcagac agttccagaa atcccagcac ccaaggaaaa acagacagca 1080ccccttgtta aatctcactc attcccatca ggttcagaac aacaaagtcc taagccttac 1140atgagaaaat ttaagacacc cttaatgatt gcggaagaaa aatacagaca acaaagggaa 1200gagcttgaga aacagagacg ggagagttct tgccatagca tcatcaaaac agaaacccag 1260caccgcagct tatcaaannt taaaaaaaaa aaaaannnag cggncgcccg 1310 37 100 PRTRattus norvegicus 37 Met Thr Arg Asp Phe Gln Lys Gln Ala Leu Ile Arg GlnGlu Lys Gln 1 5 10 15 Asn Ser Asn Lys Asp Met Arg Lys Asn Asp Met GlyLeu Gln Pro Leu 20 25 30 Pro Val Gly Lys Asp Ala His Ser Ala Pro Gly ValThr Val Ser Gly 35 40 45 Lys Asn His Lys Arg Thr Gln Ala Pro Asp Lys LysGln Arg Ile Asp 50 55 60 Val Cys Leu Glu Ser Gln Asp Phe Leu Met Lys ThrAsn Thr Ser Lys 65 70 75 80 Glu Leu Lys Met Ala Met Glu Arg Ser Phe AsnPro Val Asn Leu Ser 85 90 95 Leu Thr Val Val 100 38 774 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 38 tctagcgaac cccttcgctttttttttttt tttttttttt ttttcccccc tttcctattt 60 attaatgggg ggaagtatgtttatgtggga tttatccact tcttttagat tctcctacct 120 gttgatctgt aattattcctagtagtctct tagagttctt agaagcatgc tgttaccgct 180 aatatttcct tttggtttggatcttactta aacatattgt ttccttactc tctttttcat 240 cccagcttgt ctaactgaaaggccagaccc aacttgatct atccctttaa aacttcatgt 300 cttggcctgt tgatttctctgctccaggtg tcaccgaagg ggttcgccta gcgaacccct 360 tcgtaacagc caaggtttttgagacagagg tttcaacagc attcctggag gagacacaaa 420 ggacagatga gtcacatgaaggatgggagg agggaaggtg gctgttgata ggtattttga 480 gacactctat ttgagtcctacacaacactc ccccctcccc ccaaaccatt tttatgtcta 540 ttgacctttc ctctagtcatacagggaaat tcacagttac ctacaaagaa ccactaattg 600 taacaagtca agaggaaacttatttttgat aatgactcat tgaagatgtt ttgaaaattt 660 aaaaataagc tctgttagcagaagtctgtn ngaaaagcan gaaggaantg tttgtttatt 720 anataaataa aaggcggcgaggacaacaaa aaaaaaaaaa aaaaaagcgg ccgc 774 39 65 PRT Rattus norvegicus 39Met Ser Trp Pro Val Asp Phe Ser Ala Pro Gly Val Thr Glu Gly Val 1 5 1015 Arg Leu Ala Asn Pro Phe Val Thr Ala Lys Val Phe Glu Thr Glu Val 20 2530 Ser Thr Ala Phe Leu Glu Glu Thr Gln Arg Thr Asp Glu Ser His Glu 35 4045 Gly Trp Glu Glu Gly Arg Trp Leu Leu Ile Gly Ile Leu Arg His Ser 50 5560 Ile 65 40 1259 DNA Rattus norvegicus 40 tctagcgaac cccttcgcgaaggggttcgc cgaaggggtt cgcttcagga gttaatgtag 60 acttgactta agcatcctgatttaaccaag aatggtggca cacaacttta acccccatgc 120 tggggaagca gaggcacacttaatctgtgt gagtcccagg ccatccaggg ataccgtagt 180 agtgagaccc tgtctcacaaaacaaagaat gggaatttag ggctggtggg gctcagcatg 240 caactgtgcc tgttacctagtctggcctga gttcaattcc caagactcaa tgtatgagga 300 gagaaacgat ttctgaactcattcattgat ctccaaatgt gtggtatagg tgcccttccc 360 ttaaataaaa caaacaaacaaaaaacaaca aaaacaacaa acccccaata aatgtatatt 420 taattttaaa agactgtacttgggcatggt acttcacatc tacagttacg acattctaga 480 ggctcaggcc tgggaattgctatgaatttg aggccagtct gggttagagt gacttctcat 540 ctaggcagga ctacgtaataagtctttgcc caaaaataaa cagcaaccca aataagagca 600 acaagaattc tccctccaaatagtaacctg ggcctggaga gacagcttag caactgagtg 660 cttgccgagc catcgaggactggagtctgg attccagcac ccgtgtgaca gacaagctgg 720 gcgttcactc atgctgatgaaccccaaggc tgaggagaca ctgactcttc tctggccctg 780 ttcatgctgt ccacaggtgcccaagtagca gttaagtaga ctgtcagaca acatggctgg 840 ctttttaagc aagaacagtaactgaagaaa tacacttttg aagtactgtt aattttgctt 900 aaaacttggt agggagctggaggatggctc agtggttaag agcactgact gctcttccag 960 aggtcctgag ttcaattcccagcaaccaca tggtggctca caaccatctg taatgagctc 1020 tgatgccctc tttttggtgtgtctgaagac agcgacagtg tactcatata aaataaaata 1080 aatctttttt ttttttaaaagaaatttgtc agagatatgg caggaagggt atatttttac 1140 ctatttacct ggtgggctaatcctggtatt tttttcaaaa ttaagatact atataggagc 1200 cgcgaagggg tcgctaggccagtgtgatgg atatctgcag aattcggtta gccgaattc 1259 41 42 PRT Rattusnorvegicus 41 Met Val Ala His Asn Phe Asn Pro His Ala Gly Glu Ala GluAla His 1 5 10 15 Leu Ile Cys Val Ser Pro Arg Pro Ser Arg Asp Thr ValVal Val Arg 20 25 30 Pro Cys Leu Thr Lys Gln Arg Met Gly Ile 35 40 42777 DNA Rattus norvegicus 42 tctagcgaac cccttcgtct cctcttaaac atcttaagacaagctgttat catctacact 60 gctcttagta ctgttctttt ctaagattct tctaatatgacacattaaga ctttcttaaa 120 atgtacaact gctacgctga tctaaacatt caaagtgcacacatttcgct atgaagccac 180 gtgaccagag tcctggggac taatttctgt cttagtcagattcctattgc tatatgaaga 240 aataccatga tagtgtcaac ttttataaag aaaaagtattcctttgggaa tagtttaaag 300 gatcagaggg ttagtgcatt atcatcacag caggaagcgtggcagtggga gcccagattt 360 ctatatccag attttcatga agcatgacga gagctcctgggcctggcgcg agcttctgaa 420 acctgaaagt gacatatttc ttccaataag gccacaactactgctataag gccacatctc 480 ctaactgtgt cactatctat gagcctgtac agtctatttcttttacacca ctgcatcatc 540 taagagctga tacccgttaa gttagtcatg aaaatattcaacttctaggg ttctgttttc 600 ttctctataa aatattgaaa atgataatta atgtatactttacagaactg tatttgaagt 660 acaacttgat ggacataaat caccacagtt gggtcaaaattgtatatata tatatatata 720 tatatatata tatatatata tatcaaaaaa aaaaaaaaaaaaaaaaaaag cggccgc 777 43 46 PRT Rattus norvegicus 43 Met Ile Val SerThr Phe Ile Lys Lys Lys Tyr Ser Phe Gly Asn Ser 1 5 10 15 Leu Lys AspGln Arg Val Ser Ala Leu Ser Ser Gln Gln Glu Ala Trp 20 25 30 Gln Trp GluPro Arg Phe Leu Tyr Pro Asp Phe His Glu Ala 35 40 45 44 1378 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 44 tctagcgaac cccttcgtacatttcaccct agaaataaat agaccttcta gctctgacag 60 aaagtagtgc ttgcctaggaggagctgggc tggccagttc ctccttcttg cacacttagc 120 ctgtttgctg aaggcttgtttcaatggaaa actgaaatgg acccactaat gtctcgattc 180 ttctctcctt cactaagtctgtgaagtcat cagcgttttg tcttttgtgt gtgaataccg 240 aggagaattt cctcacccagtgccttcagg agccatgatg gctgcctcag aataagcaca 300 gatacacttg agcaactggtgcagaaaacc cgacttctaa attattaagg aacaggataa 360 ttgcttgtta caataattagaataatgtaa ttaggataat tgcttttaaa aaatcttccc 420 acctttcccc ccccaaatattaataattcc aactaaatcc tctggggccc ttccagtttc 480 cacaacggaa agagcctaacgtattctaaa gactgggcat attttttttt tccagattag 540 tgagtgttca tgagctattaagaggccaag tgttttttca agatggtgtc atttcattct 600 aacatatcta acatgcaaaggacttaaaaa aataatttgc aaaataatct gtttcaagtc 660 tatgaggaag ctgaagagcctactccggag gaaactccag aagagcctcc tagcatagag 720 gaagaagaga tagtggaggaagaggaggag gaggaggtgc ccccgcccag aggtacagcc 780 gctttgatga gttcagcattccaaagcctt ggtgctgctg gaccctactc attagccata 840 tactttcctg gaagcacagccacgaggcct ggagggtgca cactcgtaat gactggagct 900 ttgtgggcct ttcctttcccctaacgtttc ctccttcccc gcaatctgac cataaatgag 960 gagatttttt ttttctcttactacactttt tgcaatccta gtttgcaatc ctcagtgtgg 1020 ctggctttca gttcaaatgctggagaacca tgtatctgtg tggtgagagc attcattttc 1080 aagactaatt cttaaaccgcttatccccgg agacagaaac cgtggcagag ttgctatcct 1140 ctgagctggg gtggtcatgatgatcagtta ggttactaac atcttcctaa atgaatcggt 1200 gttttgtgtt gctctgttttcatttggatg acagggtgtt gttctgttta atgcgtgtgg 1260 gtttttccaa catgtccgtaaaaatatctt ttaagcacca gangtagtga agaaagctgt 1320 gcaaacagca cccgctcctgtccccaagaa awccgaggcg cccccccaaa ggtatatc 1378 45 1554 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 45 tctagcgaac cccttcgcgaaccccttcgc tgcatcctca taaagctacc tcaagacaga 60 gcgtaactgc ctcattctaggagtggactc ggggaagaca gcagacacac catcagggag 120 cccctgggta tctccagaacatggcaagcc gtggatacct gcatcacctg ctgactgcag 180 agggagcctg ggaggagtttgtatcaaagg ccaagttgcc cagggatagg gcagtggccc 240 tccacaaagc actgagggatctgacagcac tcttggccat agcagaaaga ggcagatctc 300 ggaaaggctg gaaaggcaaggagaagtttg tgaaagcatt tccttgcttg aaagcagact 360 tggaggagca catcagccagctctatgccc tagccgacca tgctgaggaa ctgcacaggg 420 gctgcaccgt ctccaacatggtggctgact ccttcagtgt tgcctccgac atcctgaaca 480 tctttggtct ctttctggcacctgagtcag cagagggaag tctggtgctc tcggcagcag 540 gcttggggct gggggtagcagctactgtga ctaatgttgc tacttcaatc atgaaggaaa 600 caagcagggt tttggatggagtcgaagctg gtcaccatgg ttcaaccgcc atggatatac 660 tggaggaagc tggcacaagtgtggctagga ttgccagcga gatccctcag gctaccagag 720 atatcaccag agacctggaagcccttgagc agcacatgaa tgccctcagt ctggtcagag 780 ccaaccctcg cctagaagaagatgccaggg ccctcatcaa tgcaggtagc atccctgccc 840 aacgggctaa acaggtgcgggccagtctga aaggaacccc tctggcaatg agcaaggaag 900 accggatccg cagtgccaccaccactgggg tcaccctctt gcgtgatgtg gggagccttg 960 tgaacgagtc gaagcagttgtacgaagggt ctgcttccga atcggcagca gcactaagga 1020 agctggctca ggagctggaggagaagctag gggagctcat gaaattctac gagacaatct 1080 gatcaggttt cagccagtcaccccatcccc aagacatgca gacatcangg gagaggatct 1140 ggacagaggt agggaccatggaggtgctgt tagaaggaga gcaagactac agtcaggtcc 1200 gagggacata gtgtggaggcctgtttgatg aacacarcag gttaraggat ggagcagtgg 1260 atcaaagtga gatccactggagcctgagac sagggaccag aggatgtgct gcaagaggga 1320 ctgggaaaat tgaaatctanactaaacatg gaaaaaaggc agtttcgaaa gactagaaaa 1380 ccctccccat ctgagccattggaaacccca caaaacacaa accagagaga aaagtgtgtg 1440 ctctctaaac aagtcgtggcccccagttcc ccagcccact cccaccctca ggggtggcat 1500 caaataaatt gtttccatttcaaaaaaaaa annaaanaaa aaaaaagcgg ccgc 1554 46 313 PRT Rattus norvegicus46 Met Ala Ser Arg Gly Tyr Leu His His Leu Leu Thr Ala Glu Gly Ala 1 510 15 Trp Glu Glu Phe Val Ser Lys Ala Lys Leu Pro Arg Asp Arg Ala Val 2025 30 Ala Leu His Lys Ala Leu Arg Asp Leu Thr Ala Leu Leu Ala Ile Ala 3540 45 Glu Arg Gly Arg Ser Arg Lys Gly Trp Lys Gly Lys Glu Lys Phe Val 5055 60 Lys Ala Phe Pro Cys Leu Lys Ala Asp Leu Glu Glu His Ile Ser Gln 6570 75 80 Leu Tyr Ala Leu Ala Asp His Ala Glu Glu Leu His Arg Gly Cys Thr85 90 95 Val Ser Asn Met Val Ala Asp Ser Phe Ser Val Ala Ser Asp Ile Leu100 105 110 Asn Ile Phe Gly Leu Phe Leu Ala Pro Glu Ser Ala Glu Gly SerLeu 115 120 125 Val Leu Ser Ala Ala Gly Leu Gly Leu Gly Val Ala Ala ThrVal Thr 130 135 140 Asn Val Ala Thr Ser Ile Met Lys Glu Thr Ser Arg ValLeu Asp Gly 145 150 155 160 Val Glu Ala Gly His His Gly Ser Thr Ala MetAsp Ile Leu Glu Glu 165 170 175 Ala Gly Thr Ser Val Ala Arg Ile Ala SerGlu Ile Pro Gln Ala Thr 180 185 190 Arg Asp Ile Thr Arg Asp Leu Glu AlaLeu Glu Gln His Met Asn Ala 195 200 205 Leu Ser Leu Val Arg Ala Asn ProArg Leu Glu Glu Asp Ala Arg Ala 210 215 220 Leu Ile Asn Ala Gly Ser IlePro Ala Gln Arg Ala Lys Gln Val Arg 225 230 235 240 Ala Ser Leu Lys GlyThr Pro Leu Ala Met Ser Lys Glu Asp Arg Ile 245 250 255 Arg Ser Ala ThrThr Thr Gly Val Thr Leu Leu Arg Asp Val Gly Ser 260 265 270 Leu Val AsnGlu Ser Lys Gln Leu Tyr Glu Gly Ser Ala Ser Glu Ser 275 280 285 Ala AlaAla Leu Arg Lys Leu Ala Gln Glu Leu Glu Glu Lys Leu Gly 290 295 300 GluLeu Met Lys Phe Tyr Glu Thr Ile 305 310 47 1142 DNA Rattus norvegicusunsure (0)...(0) n = A, T, C, or G 47 tctagcgaac cccttcggct ttttctgatttaaagtgaag aaatggccat atttgcttga 60 taatcttcag ttgtgtctct ggaactcaacaaagaacgca ttttatgaaa tatacagctg 120 tcttcggtaa agccaacttt cttacacatatttcgggaag taattaacta caatttggac 180 ttatagttac aaggttgcct tcgaaacactgctctaaatg tgtctcgtgt tggggtgcta 240 ctttgcttat gtgtaaattt cacagtaatgcaatagagaa agggtgtttg tgggtgtggc 300 ttgtgggggg gattgttttg ttgttgttgtttgagataaa gcttcattct gtagccagga 360 aagcctggaa tttactgtgt catcccaggtagcttcaaac tggtgcctat cctgcctcag 420 cctccaacgt gttgcaattg caggagtaacctaccacatc ctgcagctac agtgatctag 480 aacctccccg tcgaagcccc accaccatagaaaccaattt gcattaagtt ttagaattcc 540 caacccaact aaagtttaat aaaaaaagaaaaacaaaaca agatttaaat cattctttcc 600 ctcattcttt ttnnagatnc agggctcncctagttttnaa caaaacagtn ngcagngnng 660 ggnnccccng gnggggnttt tttncnttgngccncntngc ancccacccn cccaggcngg 720 atngggnggg gtataaaagt nttancnggcanatgnnctn ggngcanacc caagtntatc 780 aggncctnan ttnccnccca ganaactagananctntngc atagtanang ccccntgtgn 840 agatttnaaa nccncctgtn cacagananagaancttana tagaaaantc aaaatatttn 900 ggngcccaan gttncccacc ctgtagagngggnccccaaa ancngccncc aganagcnng 960 atatntgagt tntgacctnt attctttactacnacgcntt gagagaatat tntgntgggg 1020 ccctanccac atgttttgnc ccaagantgtaaanccactt naannctgng ggatatctcn 1080 ctgcanacag aagtgcccng cgggattttaaaaaaaaaaa taaaaaaaaa aaaggngccn 1140 cc 1142 48 502 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 48 tctagcgaac cccttcgtggagactgtgga agttatgtat gaataggaga gtgtgtgttg 60 tgtaacacag acagaaggacattggatcat gttgaacccg cacccccaac tatgagtgat 120 ggtatggaaa gaatgcgaacatttaaactg cgccaatgcg gcggccatct tggtggagaa 180 gttcctagcc gagctttgatgtgatttttt tgatggtaca atgcagcgag catggccacg 240 ggagctttga atccagccgacagctccgag atttgccctt ccagtgctct tgcctaccgt 300 agagaggact gctgagatgggattccttgt gacaagccta cttaccttta actgccagca 360 tttgtaaggt gcaatcttgtgtattggttt tttattttga cagttttgaa aacatgtttg 420 ntgntcttgg tgtttttccagtaaaagtaa tcacaaagga aaaaaaaatt aaaaaaaaaa 480 aaaaaaaaaa aaaagcggcc gc502 49 1426 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 49tctagcgaac cccttcgcct tcatatggtt ttacactgta tgcatctcac cgcggcccgg 60aacctttctt ctcatcccaa tcctgtttga ggggacgggg ggcagggacg gacaacccaa 120gacaagggat atttgtgctg tgggtattgc atcttatgga gggctgtagc taactgggac 180tcctgggtga ccccaacagg cctttgatcc tctgtctctc cccgcttgat ctttcttacc 240ttatgcttcc ccaagtgcag ctgagggact acacagtggc tcccgcccca ctccaaacac 300aggaaatcaa tctcagggag aggagataag aagtgaggag aagccaagat tcaaccaata 360gatggtaatt gctcctggga ccgccccccc aagcatcatt tccataggaa ggactgagtt 420tggctcctga agcccagtgg agtacctttc tctgcctgaa ttctgttgtg atccctggcc 480aagtcctctt tccagaaacc ccacctttaa aaccagctga gaaggacctt cttctctatg 540tttaataggt aactttccat agcttagctt ccctgcagtc tcccgagtgc ccagttaaaa 600ttctgccata ggtcaaaagt ggggttgaga ggtgaagtca gaggccatgc atggagctca 660gaacgtttct aaacctcctg tgattcattg agtagcccct agactctaga aggctcagat 720gccaaaaagg ktgactttat aatttcttag ggtcttctca tgggatcgkt ttcagagtgg 780gcattcacta aatgatagca agtttattaa ttgtttccca gygcctgatc tctttatttn 840cccagggctt ccaaccagag cccttggttg aaagtctccc acccaccccc caccctgaga 900cttggtggnt ttctgagatt ccccagggat ggcaaaattg gcattcttac agggagccct 960gacttctagc acgttaccta gattttttac cctgctctct ctgcctattt tactatggga 1020tcactgntct ctttggactt aaggaaccac cttgaagtag agtgaggtga ccacgtgttg 1080gtggcgaaga atataagcat tggtccttaa aagagaactt ctatgaagtc aggctgcaag 1140ctttaacatg gcacaagttg caccttactg gctgctaagt ctggatgtca accaaaggtc 1200aactctntaa ttaaagaaaa gcaagggaga aganaggtgg aagnggcttn cataaacttt 1260attcaaaatg tctaccagga atggtggtga caccaataat cccacatgtt ggatgtngag 1320gcaggaagaa tgatggtaag gggcatcctc actacataat gagttgaggc tngactaggt 1380taactntgct tnaaaaaaaa aaaaaaaaaa aaaaaaaagg ggngcc 1426 50 985 DNARattus norvegicus 50 tctagcgaac cccttcgcaa gaactcagac tgctcctgcctgacttccta ggtgtcatag 60 ctctcttctg ccgccagtat gacatcatca aggacaacgagcccaataac aacaaggaaa 120 aaaccaagag tgcatcagag accagcaccc cagagcaccagggtgggggt ctcctccgaa 180 gcaagatatg aaaccctttc agtgcttgct ctgagcagctcagaagtaga atgcgagagg 240 acctcactgt tctgacgatg attgtccaac acacatccggccctctccgt gtctcctccc 300 accaccatct tctcctatca ccgggcttac tatcttctctcctggctttc ctctttctga 360 tggcggttcc tgaagcctcc aactaacccc taactcggggagcgcctcga cagtgtttgt 420 ggctaaggct acactcagag acagagttgc agaatgagggagacccagcc cgagggacgc 480 cattgctggg aggtagactg ggtgcgaggg cccttggcacaggactcaca tctgggctgt 540 tcagcttgac ccgaaggctg tgtgtgaaag ggggaaaaagacaagattgc caggcagggc 600 tgttgttttt gtggcttcga gggacaagaa cctggctaaaaggcagcagc cctgctgttc 660 tttttctcct ctgtcctgtt tcctacctta caagaagtccatgcaaccaa ccggggctct 720 ggcacttttc ttgtttattt ccctcctggc ttccaaacaagccctctgtg gacatcatca 780 aagcatggat aaccccctct gcaggggtgg gcttcattctccgctggtcc ctgtagcctt 840 cctggacaca gggtgaaagt tgtaaaagtg gtaggagtgcagctagccac aggttctcct 900 tttcccatct cagtctgacc aaggaggctg aactaccaacccaaattcag cgaaaaaaaa 960 aaaaaaaaaa aaaaaaagcg gccgc 985 51 58 PRTRattus norvegicus 51 Met Thr Ser Ser Arg Thr Thr Ser Pro Ile Thr Thr ArgLys Lys Pro 1 5 10 15 Arg Val His Gln Arg Pro Ala Pro Gln Ser Thr ArgVal Gly Val Ser 20 25 30 Ser Glu Ala Arg Tyr Glu Thr Leu Ser Val Leu AlaLeu Ser Ser Ser 35 40 45 Glu Val Glu Cys Glu Arg Thr Ser Leu Phe 50 5552 2010 DNA Rattus norvegicus 52 tctagcgaac cccttcgcgg ggacagacatggagaaggag atggaggacc ccctggctgg 60 agcagaccaa cagaataggc aactatggctggagaaccgg gtatcagagt aatgcttgac 120 ctcgggaaac accaaatttc ttcttccgatcgcagaagta gtactcggcg aaattcacta 180 ggtaggaggc tcctcatctg ggaagaaccggtgcctgggg ggacctggct ggataggtat 240 gggggatcga ggccggtccc ctagtctccggtccccccat ggcagtcctc caactctaag 300 caccctcact ctcctgctgc tcctctgtggacaggctcac tcccagtgca agatcctccg 360 ctgcaatgcc gagtacgtct cgtccactctgagccttcgg ggagggggct caccggacac 420 gccacatgga ggcggccgtg gtgggccggcctcaggtggc ttgtgtcgcg ccctgcgctc 480 ctacgctctc tgcacgcggc gcaccgcccgcacctgccgc ggggacctcg ctttccactc 540 cgcggtgcat ggcatagagg acctgatgatccagcacaac tgctcacgcc agggtcccac 600 ggcctcgccc ccggcccggg gtcctgccctgcccggggcc ggcccagcgc ccctgacccc 660 agatccctgt gactatgaag cccggttttccaggctgcac ggtcgaaccc cgggtttctt 720 gcattgtgct tcctttggag acccccatgtgcgcagcttc cacaatcact ttcacacatg 780 ccgcgtccaa ggagcttggc ccctactagataacgacttc ctctttgtcc aagccaccag 840 ctccccggta gcatcgggag ccaacgctaccaccatccgg aagatcacta tcatatttaa 900 aaacatgcag gaatgcattg accagaaagtctaccaggct gaggtagaca atcttcctgc 960 agcctttgaa gatggttctg tcaatgggggcgaccgacct gggggctcga gtttgtccat 1020 tcaaactgct aaccttggga gccacgtggagattcgagct gcctacattg gaacaactat 1080 aatcgttcgt cagacagctg gacagctctccttctccatc agggtagcgg aggatgtggc 1140 acgggccttc tctgctgagc aggatctacagctgtgtgtt gggggatgcc ctccgagcca 1200 gcgactctct cgctcagagc gcaatcgccgtggggcgata gccatagata ctgccagaag 1260 gttgtgtaag gaagggcttc cggttgaagatgcctacttc caatcctgcg tctttgatgt 1320 ttcagtctcc ggtgacccca actttactgtggcagctcag tcagctctgg acgatgcccg 1380 agtcttcttg accgatttgg agaacttgcaccttttccca gtagatgcgg ggcctcccct 1440 ctctccagcc acctgcctag tccggcttctttcggtcctc tttgttctgt ggttttgcat 1500 tcagtaagta ggccagcaac ccgtgactagtttggaaacg gtttgaggag agaggttgat 1560 gtgagaaaac acaaagatgt gccaaaggaaacagtgggga caggagacaa cgaccttact 1620 caatcacacg aggttgcagt ccagggctgaaatgacccta gaataaagat tctgagacag 1680 ggttttgcac tccagacctt ggtatgggctccccatgaat ttccccatta gtgatttccc 1740 acttgtagtg aaattctact ctctgtacacctgatatcac tcctgcaagg ctagagattg 1800 tgagagcgct aagggccagc aaaacattaaagggctgaga tatcttaaag gcagaaacta 1860 gaaaagggga aaccatgatt atctataagaaaatcaaaag aggggtttgg gaatttagct 1920 cagtggtaga gcacttgcct agcaagcgcaaggccctggg ttcggtcccc agctcctaaa 1980 aaaaaaaaaa aaaaaaaaaa aagcggccgc2010 53 422 PRT Rattus norvegicus 53 Met Gly Asp Arg Gly Arg Ser Pro SerLeu Arg Ser Pro His Gly Ser 1 5 10 15 Pro Pro Thr Leu Ser Thr Leu ThrLeu Leu Leu Leu Leu Cys Gly Gln 20 25 30 Ala His Ser Gln Cys Lys Ile LeuArg Cys Asn Ala Glu Tyr Val Ser 35 40 45 Ser Thr Leu Ser Leu Arg Gly GlyGly Ser Pro Asp Thr Pro His Gly 50 55 60 Gly Gly Arg Gly Gly Pro Ala SerGly Gly Leu Cys Arg Ala Leu Arg 65 70 75 80 Ser Tyr Ala Leu Cys Thr ArgArg Thr Ala Arg Thr Cys Arg Gly Asp 85 90 95 Leu Ala Phe His Ser Ala ValHis Gly Ile Glu Asp Leu Met Ile Gln 100 105 110 His Asn Cys Ser Arg GlnGly Pro Thr Ala Ser Pro Pro Ala Arg Gly 115 120 125 Pro Ala Leu Pro GlyAla Gly Pro Ala Pro Leu Thr Pro Asp Pro Cys 130 135 140 Asp Tyr Glu AlaArg Phe Ser Arg Leu His Gly Arg Thr Pro Gly Phe 145 150 155 160 Leu HisCys Ala Ser Phe Gly Asp Pro His Val Arg Ser Phe His Asn 165 170 175 HisPhe His Thr Cys Arg Val Gln Gly Ala Trp Pro Leu Leu Asp Asn 180 185 190Asp Phe Leu Phe Val Gln Ala Thr Ser Ser Pro Val Ala Ser Gly Ala 195 200205 Asn Ala Thr Thr Ile Arg Lys Ile Thr Ile Ile Phe Lys Asn Met Gln 210215 220 Glu Cys Ile Asp Gln Lys Val Tyr Gln Ala Glu Val Asp Asn Leu Pro225 230 235 240 Ala Ala Phe Glu Asp Gly Ser Val Asn Gly Gly Asp Arg ProGly Gly 245 250 255 Ser Ser Leu Ser Ile Gln Thr Ala Asn Leu Gly Ser HisVal Glu Ile 260 265 270 Arg Ala Ala Tyr Ile Gly Thr Thr Ile Ile Val ArgGln Thr Ala Gly 275 280 285 Gln Leu Ser Phe Ser Ile Arg Val Ala Glu AspVal Ala Arg Ala Phe 290 295 300 Ser Ala Glu Gln Asp Leu Gln Leu Cys ValGly Gly Cys Pro Pro Ser 305 310 315 320 Gln Arg Leu Ser Arg Ser Glu ArgAsn Arg Arg Gly Ala Ile Ala Ile 325 330 335 Asp Thr Ala Arg Arg Leu CysLys Glu Gly Leu Pro Val Glu Asp Ala 340 345 350 Tyr Phe Gln Ser Cys ValPhe Asp Val Ser Val Ser Gly Asp Pro Asn 355 360 365 Phe Thr Val Ala AlaGln Ser Ala Leu Asp Asp Ala Arg Val Phe Leu 370 375 380 Thr Asp Leu GluAsn Leu His Leu Phe Pro Val Asp Ala Gly Pro Pro 385 390 395 400 Leu SerPro Ala Thr Cys Leu Val Arg Leu Leu Ser Val Leu Phe Val 405 410 415 LeuTrp Phe Cys Ile Gln 420 54 705 DNA Rattus norvegicus unsure (0)...(0) n= A, T, C, or G 54 tctagcgaac cccttcgtgg ggattaaggt tctctatagctaagcctgtc ngaatgacaa 60 cacccagaga tctcacctgg ggtggtggga gcactctctgtcttgaggga acatgtacct 120 actctctcct tccacaagag ccacatacac ttagaagttccagtgaagat ctatgtgctt 180 cagaagagag gggacttgga ggtgaaaggg ggagtgggaggggggcttga ggacctanct 240 gaaagatttt angctgaaag aacttccttg attcaaagacatatgtcagt ngacccaaca 300 atgagaatga atatgagggc caggaaaact tgtgggaatcagtctcaaga cngaaacnga 360 gaaagaaaga aaagtggnta ggactcanat tggggaacctgggtagacag gagtggcnag 420 ggaagaaagg gatcttgggt tntccacagt ttgagacacatccggngntc gaccctattc 480 ccngaagccn cannanatgt tgcttccccn tcnntnnaatgggcctggng gtcctnctcc 540 ctttncccng gacatgaaaa ngtnttctgc nnanataacccccntctttc ctcccccttn 600 antntgtccc taccnttttg tccctttttn ttttnaaaaaannaaaataa aggggnncnn 660 tnttcccttn gaaaaaaaaa aaaaaaaaaa aaaaaaccgcccncc 705 55 58 PRT Rattus norvegicus 55 Met Thr Thr Pro Arg Asp Leu ThrTrp Gly Gly Gly Ser Thr Leu Cys 1 5 10 15 Leu Glu Gly Thr Cys Thr TyrSer Leu Leu Pro Gln Glu Pro His Thr 20 25 30 Leu Arg Ser Ser Ser Glu AspLeu Cys Ala Ser Glu Glu Arg Gly Leu 35 40 45 Gly Gly Glu Arg Gly Ser GlyArg Gly Ala 50 55 56 968 DNA Rattus norvegicus unsure (0)...(0) n = A,T, C, or G 56 tctagcgaac cccttcgcga aggggttcgc ttacattcac gcttaagcatattaactgta 60 catattaact gatttagagg atactatgga ttccacatct tccctgagcatagggattga 120 tttgaaaaat gacagggttg gctgtcgacc cccatcggag gaagcaggtaaggaatcact 180 taggagaact gatctcaaca ttcttcagtt ctttctatta tttacttgtttagcctggag 240 ttaaattccc actccttgtg agcacttcta atttgaaaat ccactttcttcaatattttc 300 gaaatttaaa actgatggat gacgtgacaa aacttccacg agttaagaattctccacctc 360 tgatctcatc gcagcagggc acaatccaag gcatgtgaat tgacttccaggtttatgtga 420 catataaatg aattctgtct ctagatttgg atcccattct cctaaatatctcaccatgca 480 tgtgcagata ttctaaagtc taaaaatatc tgatattgca aacttttctggtcaaaacat 540 tttggatgag ccatttaaca gccaaggtat ttgagacaga ggtttcaacagcattcctgg 600 aggagacaca aaggacagat gagtcacatg aaggatggga ggagggaaggtggctgttga 660 taggtatttt gagacactct atttgagtcc tacacaacac tcccccctccccccctcccc 720 ccaaaccatt tttatgtcta ttgacctttc ctctagtcat acagggacattcacagttac 780 ctacaaagaa ccagaattgt aacaagtcaa gaggaaactt atttttgataatgactcatt 840 gaagatgttt tgaaaattta aaaataagct cttgtaagca gaagtctgtgagaaaagcaa 900 gaaggaattg tttgtttatt aaataaataa aaggcnnann nnaaaaaaaaaaaaaaaaan 960 gcggccgc 968 57 52 PRT Rattus norvegicus 57 Met Asp SerThr Ser Ser Leu Ser Ile Gly Ile Asp Leu Lys Asn Asp 1 5 10 15 Arg ValGly Cys Arg Pro Pro Ser Glu Glu Ala Gly Lys Glu Ser Leu 20 25 30 Arg ArgThr Asp Leu Asn Ile Leu Gln Phe Phe Leu Leu Phe Thr Cys 35 40 45 Leu AlaTrp Ser 50 58 1183 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C,or G 58 tctagcgaac cccttcggca gacagcatcc ctcccaaggc tactcagggtttaaaccctg 60 cttctgaagt gacatgtcct gcaaagaaag tccccacgtg ggtgtttccaccaccactgt 120 cagctctgta gctgtgcaag ctggggactc caagatcgtg atagccgttgtcaagtgtgg 180 caaatgggtg cggctccaac tggctgaggc acagcccaat ctcctagaaattgggagcag 240 tcaagatgaa accagaaaac tgcttcacga tcacgagctc cttctggccaagcttaaggc 300 cttggaagat cgtgtgtggg gactcttaca ggaagcagac aggacggctgaagcaaacaa 360 ggagcaaagt gaggtgtcga tgccatggcc agactctggg cgaagcatgggccaccctgg 420 tcttcatgct tgaaagaaga agggagctcc tcggactgac atctgagttttttcaaagcg 480 ccttggagtt tgctataaaa atagaccaag ctgaagattt tctgcagaatcctcacgagt 540 ttgagagtgc cgaagcctta cagtcacttc ttctgcttca tgaccgacacgccaaagaac 600 tcttagaacg atctctagtc cttttaaaca aaagccaaca actcactgacttcatagaaa 660 aattcaagtg tgatggatct cctgtgaatt ctgagctcat ccagggagctcagagcagtt 720 gtctgaagat cgacagcctc cttgaacttc tgcaagacag gagaaggcagctggacaagc 780 acttgcagca acagaggcag gagttgtctc aggttctgca gttatgtctgtgggaccaac 840 aagaaagcca ggtttcttgt tggtttcaga aaacaataag agatctgcaggaacagagtc 900 tgggttcatc cctttcagac aacaaagagt taatccgtaa gcacgaggacctgccatcaa 960 agcaaagagt ccctgcagtt taggaattga acagaacagt ttcctgattgaatgatcttg 1020 gcgcctyytt ancggntgca gatggtgggg cttcctctgg nttctcatcctcttccacta 1080 atctggattt ttgttcccct ggtgtgccac atcactttaa tttgaaagaaaaaaaataaa 1140 ttgggccgga aaaaaaaaaa aaaaaaaaaa aarrrscggc cnc 1183 59245 PRT Rattus norvegicus 59 Met Lys Pro Glu Asn Cys Phe Thr Ile Thr SerSer Phe Trp Pro Ser 1 5 10 15 Leu Arg Pro Trp Lys Ile Val Cys Gly AspSer Tyr Arg Lys Gln Thr 20 25 30 Gly Arg Leu Lys Gln Thr Arg Ser Lys ValArg Cys Arg Cys His Gly 35 40 45 Gln Thr Leu Gly Glu Ala Trp Ala Thr LeuVal Phe Met Leu Glu Arg 50 55 60 Arg Arg Glu Leu Leu Gly Leu Thr Ser GluPhe Phe Gln Ser Ala Leu 65 70 75 80 Glu Phe Ala Ile Lys Ile Asp Gln AlaGlu Asp Phe Leu Gln Asn Pro 85 90 95 His Glu Phe Glu Ser Ala Glu Ala LeuGln Ser Leu Leu Leu Leu His 100 105 110 Asp Arg His Ala Lys Glu Leu LeuGlu Arg Ser Leu Val Leu Leu Asn 115 120 125 Lys Ser Gln Gln Leu Thr AspPhe Ile Glu Lys Phe Lys Cys Asp Gly 130 135 140 Ser Pro Val Asn Ser GluLeu Ile Gln Gly Ala Gln Ser Ser Cys Leu 145 150 155 160 Lys Ile Asp SerLeu Leu Glu Leu Leu Gln Asp Arg Arg Arg Gln Leu 165 170 175 Asp Lys HisLeu Gln Gln Gln Arg Gln Glu Leu Ser Gln Val Leu Gln 180 185 190 Leu CysLeu Trp Asp Gln Gln Glu Ser Gln Val Ser Cys Trp Phe Gln 195 200 205 LysThr Ile Arg Asp Leu Gln Glu Gln Ser Leu Gly Ser Ser Leu Ser 210 215 220Asp Asn Lys Glu Leu Ile Arg Lys His Glu Asp Leu Pro Ser Lys Gln 225 230235 240 Arg Val Pro Ala Val 245 60 1051 DNA Rattus norvegicus 60tctagcgaac cccttcgcgc aagatggccg cttcccagac cgctccgcgg catcttcaag 60atgcgcgaga agaacgtgca atctcgcgag atcaggctcg ctcgcgggca gtctgctcgc 120agcctaccct tcctaggagt tggaggaggg aaagctagat tcgattaaga gcaaaaaatt 180gttccagcag cagagcagct gtccaaggaa gtatccaaag gaactgcacc tcagtaaact 240cctggcaagt cttaggatat gacaaagggc acaggatgca ttatgagaaa ggaaggctaa 300ggttttcaag aacacagatt tacatcaaac ttgcgttctg aattaatctt tgagaatact 360ggactgtgag ctagacattg agtaagaggt ttgttatatc aagaatgtga tctaaaaaaa 420aaacattcat atcttcctcc cacaagagga tattttgaaa ctgtgggtca aagtcagact 480acaggagagc cctcaaatat gccaaatgtg acagacagca ggattttgaa aatatagtgg 540gagtatgtga agatgttcca gtcaaagaga cattgtttcc aaaggaaaga aagtccagtc 600gcctcacagg aattgtgtat tccctggtag taatgcaaat ggaccacata tggctttctt 660ctttaaagag aatacctaat tttagctaca gagtaaaatg ctgatgatac aaaccgtgac 720aagtggaggg acaagaaagt aaatggactg atggtgccat tgtggactgg gagggtaaaa 780gctgtacatt tgtgaacaaa aagatttcct tgttatggtc agccatgatt ctaactgcta 840aatggaggca gtaacaacat gacctaaaga gtaaacatcc agagatggaa tgttctcaat 900gtctgaaaag gagcagatat ctggtgtatg tgaatgtatg ctagagattt tttacaagcc 960tgtggtgaat tagtaattgt attttatttt gaaagttaaa caggtaatta gaaaccccaa 1020aaaaaaaaaa aataaaaaaa aagcggccgc c 1051 61 576 DNA Rattus norvegicusunsure (0)...(0) n = A, T, C, or G 61 tctagcgaac cccttcgctg aaaccaccgttcacacggga aacctgggtt aggcttttgt 60 cctcagtgac acagaggatg tagtccacagctaggtagaa atgtcaggtt cccaacacta 120 ctccagctgt gactttgatg cttgggggatggggtcgcag gctattttct ctgctttaac 180 agttcataga atttaacaga taagagttagtgtctttcat gtggcctcac tctggagtta 240 tgagaacata cacacggttt acagcttttcaatatncctt tccctggcca tcaagtattt 300 tgaaagtgtg ccacctttta acctttgcgctttatttttt tttctttttt taaagntgaa 360 ggtgataatt cttctatata tgatgaaactcaatgtctac tgaaataagt gtaaccttag 420 ctatncacgt ttatntttta aaaccacgctatggagatat taccccgagt tctgtcnttt 480 ngcaagattt acagnacctt cccncccccccttttagcat tnaataaaaa natattgggg 540 agcncnntna aaaaaaaaaa aatnaanaaaagcggc 576 62 587 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, orG 62 tctagcgaac cccttcgcgt gatctgatcc gagctgagac ttggggagct ctggctccgt60 gttggctgca gcatccccca tggtcttgtc tgaggtgtcc tgtgactcga ctcttcagaa 120ctcaatgaag tagatgactt gactacaatg tggaaacatc atgacagaaa gtgtggtttg 180taccggggcc gtcagcactg taaaggaagt ctgggaagaa agaataaaga aacatcatga 240agatgtgaaa cgagagaagg aatttcagca aaagctagtg cggatctggg aagaccgagt 300gagtttaact aagctgaaag agaaggtgac cagggaagat ggaagaatca ttctaaggat 360agagaaagag gaatggaaga ctctcccttc ttccttactg aaactgaatc agctacagga 420gtggcaactt cataggaccg gattgttgaa aattcctgaa ttcattggaa gattccagca 480tctcattggt ctagacttat ctcggaacac aatttcagag atccccccga ggcattggac 540tgntcactta gacttcaagg aactgattct tagctacaca aaatcaa 587 63 142 PRTRattus norvegicus UNSURE (0)...(0) Xaa = any amino acid 63 Met Thr GluSer Val Val Cys Thr Gly Ala Val Ser Thr Val Lys Glu 1 5 10 15 Val TrpGlu Glu Arg Ile Lys Lys His His Glu Asp Val Lys Arg Glu 20 25 30 Lys GluPhe Gln Gln Lys Leu Val Arg Ile Trp Glu Asp Arg Val Ser 35 40 45 Leu ThrLys Leu Lys Glu Lys Val Thr Arg Glu Asp Gly Arg Ile Ile 50 55 60 Leu ArgIle Glu Lys Glu Glu Trp Lys Thr Leu Pro Ser Ser Leu Leu 65 70 75 80 LysLeu Asn Gln Leu Gln Glu Trp Gln Leu His Arg Thr Gly Leu Leu 85 90 95 LysIle Pro Glu Phe Ile Gly Arg Phe Gln His Leu Ile Gly Leu Asp 100 105 110Leu Ser Arg Asn Thr Ile Ser Glu Ile Pro Pro Arg His Trp Thr Xaa 115 120125 His Leu Asp Phe Lys Glu Leu Ile Leu Ser Tyr Thr Lys Ser 130 135 14064 819 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C, or G 64tctagcgaac cccttcggtt ctgttggcta cacagctgca gagccatggc tgaccgttca 60ctgtcagggg cacatgttac actaagcttc atgacagtga tgtaataatg ttacacattt 120gtcttgtagt tatgtattga agtttctgtc ctgttttgtg taaaaatgta tccactcttg 180tatatattta gacttgaaac taccacacaa atattggaac ggtttgcttt atgaagttaa 240aagtatcctt ccgaatggaa ctaacttgct ttgtgctcag acatatacta tgctgatgta 300ttttgcaata tactatctta aattaaatct ggtcactttg ttgccttttt aaaaagtgtg 360gtatttcaag tagagttatt ttcctgaaat atatttgcaa actcaagctg ctttataatc 420aaggaatatt tttattgatt gaagaaaatg actgctgcaa ttcaaaagtg aacttatttt 480attatataga tgatttctta aaagctattt ataccatgat acaaaatcat gtagtgatcc 540tgggagtctg tagttcttcc tgttaataac attcaacact gtatgctaga ggcagcaatg 600ccaacactga agttattttg ggtgaaaacc gtcgttctgn cctgtttagc tggggattat 660taaatccata taatgtatgt gcttatgtat gctacatgtg caagttaggt gtttcctttg 720tgttctgctt attaaatgtc attcagattc acttcctgaa ttctaataaa gagggaagct 780attggaaaaa ataaaaaaaa aaaaaaaaaa gcggccgcc 819 65 1648 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 65 tctagcgaac cccttcggtggcgcacgccg gtaggatttg ccacgcaaat gctggaatta 60 aagacatgca gcagcagcgccctgtggttt tggtttttta tttgattgct tatttttatc 120 taatttttaa ttttttgtgtatgaacgttt tatctgcatt tatgtctctg taccacattc 180 gtgcctggtg ctatggaggccaaaaaagga ttttaggccc gagattgtag ttatagatgg 240 ttgtgggctg ccaatctgagtgctgaaaat taaacctggg tactctgaaa gaccagccag 300 tgctcttaac tatcaggccacctctccagc actattttat tttattttat ttgtggagat 360 agggtctctc tctctgtatcctagtctaac ttaaaacata aagaatattc tgtatcagta 420 tccttgagta ctaggattctaggcacctgt cattatgcct agatttttaa cagtgtgtgt 480 taattctaca taaaaatgaatttcattatt acattttcac acttgtgaag aatatacttt 540 gatcatattc ccttctcctgatactttttc ctatccttcc tccccactcc attagttccc 600 ttcttctttt cagagtctaccttctacttt ttactttgat ttttttcccc ccacattctg 660 tggttgagag aatgcatattacagttgtat ttctgaatct ggctaggtac attcacttaa 720 cataattaat gatcctgggcgagcgaaggg gttcncctan cnaacccctt cggttcaata 780 ccatttcaga gatgggcatttccctcaatg aaatacacaa gtaaacattc cgacattgtc 840 tttaggagtg tttgttaaaaaaaaaaaaaa aaaaaaccan ancccaaaan caaaaaaaaa 900 aaagctttgc accttgcaaaagtggtcctg gcgtgggtag attgctgtta atcctttatc 960 aataacgttc tatagagaatatataaatat atatataatt atatctccta gtccctgcct 1020 cttaagagcc gaaaatgcatgggtgttgta gacattcggt tgcactaaat tcctctctga 1080 attttggctg ctgaagccgttcatttagca actgtttata ggtggttgat gaatggttcc 1140 ttatctccat ttcttcctatgtagcttaag ccgcttcctt cacagaatct aataatctcg 1200 tctaggccat tagccctgccctttcttaac attcttgtat ttgttgaatt tggcctcctc 1260 gaaagcaata gcaactgggtggcccaccca agttttaacg cccctgattc catctatggc 1320 atttgtacca aatataagttggatgcattt attttagaca caaagcttta ttttttcgac 1380 atcgtgtttc aagaaaaaaaacaaatagaa taacaataac tatgactttg aggccaatca 1440 tttttaggtg tgtgtttgaagcatagaacg tctnttaaac tctcaatggt tccttcaaat 1500 gatgagttag tatgtaacgtaaatagcagt ttctctctct ctctctctct ttttattttt 1560 tccanataga gcactatgtaaatttagcat atcaataata caggaactat ccnccaaaaa 1620 aaaaaaaaaa aaaaaaaaaagcggccgc 1648 66 782 DNA Rattus norvegicus unsure (0)...(0) n = A, T, C,or G 66 tctagcgaac cccttcgtag aactaggagc cagtgttgac cacggtcggtggctggatac 60 cccactgcat gctgcagcaa ggcagtccag tgtggaggtc atcaatctgctcactgagta 120 tggggctaac ctgaaactca gaaactcgca gggcaaaagt gctcttgagctcgctgctcc 180 caaaagtagt gtggagcagg cactcctgct ccatgaaggt ccacctgctctttctcagct 240 ctgccgcttg tgtgtccgga agtgcttggg ccgcacatgt catcaagccatctacgcact 300 aggtctgcca gaacccctgg aaaaattcct cttataccaa tagttggaaacatgttgcct 360 gctgtaggac acttaatata cacattcagt ggcttaaccc actatcctaaaaatctgctt 420 acctaattag aataaagcct tcataaatcc aaatacttgc gttgaacaaactcctggtta 480 ggttaatggn tgccaagaga taaccagaaa cctttcaagt ttttaactcttggtaattta 540 aaatcaaact gaaatagatg gaaaataata atctattttt ggataattcaaggacccttc 600 agtatctggg gctggggtcc gcattttgna tactggatag acacacacacaggtaggata 660 nggtaaatna actacttaaa gaatggcctg ggatttaagt cctccagatattttttaggt 720 ngnggtttcc taaaataaaa ttctggagtg ccaaaaaaaa aaaaaaaaaaaaaaagcggg 780 cc 782 67 49 PRT Rattus norvegicus 67 Met Ser Ser Ser HisLeu Arg Thr Arg Ser Ala Arg Thr Pro Gly Lys 1 5 10 15 Ile Pro Leu IlePro Ile Val Gly Asn Met Leu Pro Ala Val Gly His 20 25 30 Leu Ile Tyr ThrPhe Ser Gly Leu Thr His Tyr Pro Lys Asn Leu Leu 35 40 45 Thr 68 538 DNARattus norvegicus unsure (0)...(0) n = A, T, C, or G 68 gtctagcgaaccccttcggg aaacttcaac aaaggtacca gcaactacag cgccttgtcc 60 acccagatttcttcagccaa aagtctcaga ctgagaaacg gttctcggag aagcattcga 120 ccctggtgaatgatgcctac aagactcttc aggcccccgt gagcagagga ctatatcttc 180 taaagctccaaggaatagaa attcctgaag ggacagatta tagaacagac agtcagttcc 240 ttgtggaaatcatggaaatc aatgaaaaac tcgcagacgc caaaagtgag gcagccatgg 300 aagaggtagaagccactgtc agagctaaac agaaagaatt tacggacaat ataaacagag 360 cttttgaacaaggtgatttt gaaaaagcca aggaacttct tacaaaaatg agatactttt 420 caaacatagaagaaaagatc aagttaagca agaaccctct ctagttgcta acttaaaggt 480 ttaaaaataaactttgtatt tcttcannnn nnannnnnan nntnnnnnag cggccgcc 538 69 70 PRTRattus norvegicus 69 Met Glu Ile Asn Glu Lys Leu Ala Asp Ala Lys Ser GluAla Ala Met 1 5 10 15 Glu Glu Val Glu Ala Thr Val Arg Ala Lys Gln LysGlu Phe Thr Asp 20 25 30 Asn Ile Asn Arg Ala Phe Glu Gln Gly Asp Phe GluLys Ala Lys Glu 35 40 45 Leu Leu Thr Lys Met Arg Tyr Phe Ser Asn Ile GluGlu Lys Ile Lys 50 55 60 Leu Ser Lys Asn Pro Leu 65 70 70 805 DNA Rattusnorvegicus 70 tctagcgaac cccttcgcga aggggttcgc ttcttaccct gtggagaaaggggcaggagg 60 aacctcctgt gttaggagga agctggagct taccactgtg agaggacagatgtggactga 120 gaattttctt agtgctcagt ggcacttccc aaggactccc ctccccttgtgctctgtgcg 180 gtttttagga cagctaagat gactgccacc tgttgtggca ggcccgatttgtcttgttct 240 ccccttactg taccccgata taatctctgt tgatcaacag gactaccccaagaatccaca 300 tgttctcccc cgtaaccagg cagctgtctg gttcatgcct tcttcccttcaaacccaacc 360 cagcgccctt gttagtgaag aggtggtcca tggactgatg acaagttattagcactggat 420 gctgtttcca tagtgacaag cctatacctc ttcccaccct ttagtgcgcagtgggctgct 480 gcttcagtat cctcccagct cagttttatt agatcaaagc tgcccttgggcaccatgttg 540 gccacctcaa tcaccagcca aaatggtcgc tttgtccacc agaggtcaagccatctttct 600 ggcgctgtag ttcccagctc cttctaggga acaggaagtt gatattgccatgggggaggt 660 ggcggggtgt ggccgtcacc tcaatagttt tactgtaaaa gggaaatttgaacaagaaca 720 acaacaaaaa aaaaaaaaaa acaaagaaaa aaataaaaaa ctttaaaagttgaaaaaaaa 780 aaaaaaaaaa aaaaaaagcg gccgc 805 71 1407 DNA Rattusnorvegicus unsure (0)...(0) n = A, T, C, or G 71 tctagcgaac cccttcgctgggacccgcaa ctaccaactg ccgcctggat cctaggtgag 60 ctgtgggctc tgacagcgctgtggctaaca tggcacccaa aaagaagact ctcaagaaga 120 acaaacccga gatcaatgagatgaccatca tcgtggaaga cagcccccta aacaagctga 180 atgctctaaa tgggctcctggggggagaaa acagccttag ctgtgtttct ttcgaactaa 240 cagacacttc ttatggtcccaacctcctgg aaggtttaag taaaatgcgt caagagagct 300 ttctatgtga cttggtcatcggtccaaaac caagtccttt gatgtccata agtcaagtga 360 tggcttcctg cagcgagtcttctataatat ccttaaaacg atccatcgac aaaaagggta 420 gacctcaatg atatcgnccctttagggcta ccaccgtgat agcatatgca tacacnggaa 480 agctgccctt tctttatacacaataaggaa gcatcatttc tgctgctgtg tacctccaga 540 tccacactct tgtgaagatgtgcagcgact ttctgatccg agagatcagt gttgagaact 600 gcatgtatgt tgttaacatggctgaaacat actgcttgaa aaatgcgaaa gcaacggccc 660 agaaatttat ccgggataacttcattgaat ttgccgactc cgaacaattt atgaagctga 720 cgtttgaaca gattaatgagcttctcatag atgatgactt gcagttgcct tctgagctgg 780 tagcattcca gattgcaatgaaatggatag aattcaacca aaagagagtg aagcacgctg 840 cggatctttt aagcaatattcgctttggta ccatctctgc acaagacctg gtcaattacg 900 ttcaaaccgt accgagaatgatgcaagacg ctgattgtca taaactgctt gtggatgcta 960 tgaactacca cttactaccttatcatcaaa acacgttgca atctaggcgg acaagaatta 1020 gaggcggctg ccgggttctgatcactgtcg ggggacgccc tggcctgact gagaagtccc 1080 ttagtagaga cgtttatatagagaccctga aaatggatgg agcaagctta cagaaatgcc 1140 agccaagagt ttcaatcagtgtgtggctgt gatggatgga ttcctttatg tagcaggtgg 1200 tgaggaccag aatgatgcgagaaaccaagc caagcatgca gtcagcaatt tctgcaggta 1260 ccgatccccg cttcaacacgtggatccacc tgggcagcat gaaccagaag cgcacgcact 1320 tcagcctgag cgtgttcaacgggctcctgt acgccggtgg ngggcnccag tgnganggat 1380 atctgcagaa ttcggctagccgaattc 1407 72 113 PRT Rattus norvegicus 72 Met Ala Pro Lys Lys Lys ThrLeu Lys Lys Asn Lys Pro Glu Ile Asn 1 5 10 15 Glu Met Thr Ile Ile ValGlu Asp Ser Pro Leu Asn Lys Leu Asn Ala 20 25 30 Leu Asn Gly Leu Leu GlyGly Glu Asn Ser Leu Ser Cys Val Ser Phe 35 40 45 Glu Leu Thr Asp Thr SerTyr Gly Pro Asn Leu Leu Glu Gly Leu Ser 50 55 60 Lys Met Arg Gln Glu SerPhe Leu Cys Asp Leu Val Ile Gly Pro Lys 65 70 75 80 Pro Ser Pro Leu MetSer Ile Ser Gln Val Met Ala Ser Cys Ser Glu 85 90 95 Ser Ser Ile Ile SerLeu Lys Arg Ser Ile Asp Lys Lys Gly Arg Pro 100 105 110 Gln 73 2004 DNARattus norvegicus unsure (0)...(0) n = A, T, C, or G 73 tctagcgaaccccttcggac actgccagca tagacagcag cccctgctac tgtcccacca 60 ctgtaccccagagccccgac tagcagtatg ccgggagcgc cagggcctgg gcctgaggtg 120 gctgcagcctttgaggaacg gttgagtcag gcactacagg aactgcaggc agtggctgaa 180 gcaggccggtcagcggtgac ccaggcagct gatgcagccc tagccactgt agagccagtg 240 gctcaggcatctgaagagct tcgggccgag acagcagccc tgagccggcg gctggatgcc 300 ctgaccaggcaggtggaggt gctgagccta cggctgggtg ttccactcgt gccggacctg 360 gagtccgagctagagcccag cgagctgttg ctggctgctg ccgaccctga ggccctcttc 420 caggcaagctgaggatgctg ggacccccgt ggccacccgc ctgcctttag cacccgccgc 480 agctcttctgcgggcccctc tcgaagcagc agtctcatgg agcccgatcc agcagagccc 540 ccctctgccacagtggaagc agctaatgga acagagcaga ctctggacaa agtgaacaaa 600 ggcccagaggggcggagccc cctgagtgca gaggagctga tggccattga ggacgaagga 660 atcctggacaagatgctgga ccaggctacg aactttgaag agcggaagct catccgggct 720 gcgctccgtgagctccgaca aagaaagaga gaccagaggg acaaggaacg agaacggcgg 780 ctacgagaggcacgggcccg gccaggcgag agccgaagca atatggctac tacagagacc 840 accaccaggcacaagccaga gggcggctga tggctcggcg gtcagcacag ttaccaaaac 900 tgagcgggtcgtccactcca atgacggcac gcagactgcg cgcaccacca cagtggagtc 960 gagtttcgtgaggcgctcgg agaatggcag cagcaagcaa gcagcagcac cacggtccaa 1020 accaagaccttttcctcttc ctcttcctca tccaaaaaaa tgggcagtat cttcgaccga 1080 gaggaccaaaccagctcacg ttctggcagc ctggcggccc tcgaaaaacg ccaggcagag 1140 aagaagaaagagctcatgaa ggcacagagt ctgcccaaga cctaagcgtc ccaagcacgc 1200 aaggccatgattgagaaact agagaaggaa ggctcttcgg gcagtcctgg cacaccccgt 1260 acagcggtacagcgttctac cagcttcgga gtccccaacg ccaacagcat caagcagatg 1320 ttgctggactggtgccgagc caagacccgt ggctacgagc acgtggacat ccagaacttc 1380 tctccagctggagtgatggg atggctttct gtgccctggt gcacaatttc ttccctgagg 1440 cttttgactatggacagctt agcccacaaa accggcgcca gaactttgaa atggccttct 1500 catctgctgagacccatgcg gactgcccgc agctcctgga tacagaggac atggtgcggc 1560 ttcgagagcctgactggaag tgcgtgtaca cgtacatcca ggagttctac cgctgtctgg 1620 tccagaaggggctggtaaaa accaaaaagt cctaacccct gcttggggcc ccacggatgc 1680 tggtggactgtgtacccttg gtggaggtgg aggacatgat gatcatgggc aaaaagccag 1740 accctaagtgcgtcttcacc tacgtgcaat cgctgtacaa ccacctgcgg cgccatgagc 1800 tgcgcctgcgcggcaagaat gtctagccac tgctcacacc gcctgcgctg caggctgctg 1860 tcccacgcccccaacaccgg nccctncagt gngcctgcca ctgntgcccg tntgtcgaaa 1920 cacctntccccttgtcacac gcagngnttt gataaattat ttgntttnaa caaaaaaaaa 1980 aaaaaaaaaaaaaaaagcgg ccgc 2004 74 114 PRT Rattus norvegicus 74 Met Pro Gly Ala ProGly Pro Gly Pro Glu Val Ala Ala Ala Phe Glu 1 5 10 15 Glu Arg Leu SerGln Ala Leu Gln Glu Leu Gln Ala Val Ala Glu Ala 20 25 30 Gly Arg Ser AlaVal Thr Gln Ala Ala Asp Ala Ala Leu Ala Thr Val 35 40 45 Glu Pro Val AlaGln Ala Ser Glu Glu Leu Arg Ala Glu Thr Ala Ala 50 55 60 Leu Ser Arg ArgLeu Asp Ala Leu Thr Arg Gln Val Glu Val Leu Ser 65 70 75 80 Leu Arg LeuGly Val Pro Leu Val Pro Asp Leu Glu Ser Glu Leu Glu 85 90 95 Pro Ser GluLeu Leu Leu Ala Ala Ala Asp Pro Glu Ala Leu Phe Gln 100 105 110 Ala Ser75 881 DNA Rattus norvegicus 75 tctagcgaac cccttcgctc cagggcgtttgcctcctgct gacttgctct tcaccattag 60 acaagcctga cgtcaagacc ccaatggctaacgaagctaa cccttgccca tgtgacattg 120 gtcacaggct agactatggt ggcatgggccaggaagttca ggttgagcac atcaaggcat 180 atgtcacccg gtcccctgtg gatgcaggcaaagctgtgat tgttgtccag gatatatttg 240 gctggcagct gtccaacacc aggtatatggctgacatgat tgctggaaat ggatacacaa 300 ctattgccca gacttctttg tgggtcaagagccatgggac ccggctggtg attggtccac 360 cttccctgag tggttgaaat caagaaatgccagaaaaatc aaccgagagg ttgatgctgt 420 cttgaggtat ctgaaacaac agtgtcatgcccagaagatt ggcattgtgg gcttctgctg 480 ggggggtatt gtggtgcacc acgtgatgacgacatatcca gaagtcagag cgggggtgtc 540 tgtctatggt atcatcagag attctgaagatgtttataat ttgaagaacc caacgttgtt 600 tatctttgca gaaaatgatg ctgtgattccacttgagcag gtttctatac tgatccagaa 660 gcttaaagaa cactgcatag ttaattaccaagttaagaca ttttctgggc aaactcatgg 720 ctttgtgcat cggaagagag aagactgctcccctgcagac aaaccctaca ttgaggaagc 780 gaggaggaat ctcatcgaat ggctgaacaagtatatttaa cagcactcaa gcacaaattt 840 tgaataatta aattgacccg aataattaaattgacccgaa t 881 76 97 PRT Rattus norvegicus 76 Met Ala Asn Glu Ala AsnPro Cys Pro Cys Asp Ile Gly His Arg Leu 1 5 10 15 Asp Tyr Gly Gly MetGly Gln Glu Val Gln Val Glu His Ile Lys Ala 20 25 30 Tyr Val Thr Arg SerPro Val Asp Ala Gly Lys Ala Val Ile Val Val 35 40 45 Gln Asp Ile Phe GlyTrp Gln Leu Ser Asn Thr Arg Tyr Met Ala Asp 50 55 60 Met Ile Ala Gly AsnGly Tyr Thr Thr Ile Ala Gln Thr Ser Leu Trp 65 70 75 80 Val Lys Ser HisGly Thr Arg Leu Val Ile Gly Pro Pro Ser Leu Ser 85 90 95 Gly 77 25 DNAArtificial Sequence Primer specific for vector to produce “Driver DNA”.77 cgtatgttgt gtggaattgt gagcg 25 78 25 DNA Artificial Sequence Primerspecific for vector to produce “Driver DNA”. 78 gatgtgctgc aaggcgattaagttg 25 79 28 DNA Artificial Sequence Oligos corresponding topolylinker sequence. 79 gccgccagtg tgctggaatt cggctagc 28 80 28 DNAArtificial Sequence Oligos corresponding to polylinker sequence. 80cgaattctgc agatatccat cacactgg 28 81 25 DNA Artificial Sequence Oligoscorresponding to polylinker sequence. 81 ctagagggcc caattcgccc tatag 2582 25 DNA Artificial Sequence Oligos corresponding to polylinkersequence. 82 tgagtcgtat tacaattcac tggcc 25 83 20 DNA ArtificialSequence Oligos corresponding to polylinker sequence. 83 gctcggatccactagtaacg 20 84 18 DNA Artificial Sequence Oligos corresponding topolylinker sequence. 84 tttttttttt tttttttt 18

1. An isolated nucleic acid molecule comprising a poly- oroligonucleotide selected from the group consisting of: (a) apolynucleotide encoding a polypeptide having at least about 80% sequenceidentity with any amino acid sequence selected from the group consistingof: amino acids 1 to 193 of SEQ ID NO: 4, amino acids 1 to 236 of SEQ IDNO:6, amino acids 1 to 61 of SEQ ID NO: 8, amino acids 1 to 92 of SEQ IDNO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36 of SEQID NO:16, amino acids 1 to 83 of SEQ ID NO: 18, amino acids 1 to 82 ofSEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1 to 170of SEQ ID NO:24, amino acids −26 to 233 of FIG. 13 (amino acids 1 to 259of SEQ ID NO: 26), amino acids 1 to 30 of SEQ ID NO:28, amino acids 1 to30 of SEQ ID NO:35, amino acids 1 to 100 of SEQ ID NO:37, amino acids 1to 65 of SEQ ID NO:39, amino acids 1 to 46 of SEQ ID NO:43, amino acids1 to 313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, aminoacids −35 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO: 53),amino acids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ IDNO:57, amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQID NO:63, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 ofSEQ ID NO:69, amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1to 97 of SEQ ID NO:76; or a transmembrane domain (membrane spanningsegment/region) deleted or inactivated variant thereof; (b) apolynucleotide encoding a polypeptide of amino acids 1 to 233 of SEQ IDNO: 26, or amino acids 1 to 387 of SEQ ID NO: 53; (c) a polynucleotideencoding amino acids 1 to 203 of SEQ ID NO: 2, amino acids 1 to 193 ofSEQ ID NO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61of SEQ ID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to92 of SEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1to 36 of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids1 to 82 of SEQ ID NO:20, amino acids 1 to 46 2 of SEQ ID NO:22, aminoacids 1 to 170 of SEQ ID NO:24, a mino acids −26 to 233 of FIG. 13(amino acids 1 to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ IDNO:28, amino acids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQID NO: 33, amino acids 1 to 30 of SEQ ID NO:35, amino acids 1 to 100 ofSEQ ID NO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42of SEQ ID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids−35 to 387 of FIG. 29 (amino acids 1 to 422 Of SEQ ID NO:53), aminoacids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57,amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ IDNO:63, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQID NO:69, amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1 to114 of SEQ ID NO:74, and amino acids 1 to 97 of SEQ ID NO:76; or atransmembrane domain (membrane spanning segment/region) deleted orinactivated variant thereof. (d) a polynucleotide hybridizing understringent conditions with the complement of the coding region of SEQ IDNO: 1, wherein said polynucleotide encodes a polypeptide having at leastone biological activity of the polypeptide encoded by clone P00184_D11(SEQ ID NO: 1), a polynucleotide hybridizing under stringent conditionswith the complement of the coding region of SEQ ID NO: 3, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00185_D11 (SEQ ID NO: 3);a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 5, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00188_D12 (SEQ ID NO: 5),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 7, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 9, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_G01 (SEQ ID NO: 9),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 11, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_G05 (SEQ ID NO: 11),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 13, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00194_H10 (SEQ ID NO:13),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 15, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00199_D08 (SEQ ID NO: 15),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 17, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00203_D04 (SEQ ID NO: 17),a polynucleotide hybridizing under stringent conditions with thecomplement of the codin region of SEQ ID NO: 19, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00203_E06 (SEQ ID NO: 19),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 21, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00209_F06 (SEQ ID NO: 21),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 23, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00219_D02 (SEQ ID NO: 23),a polynucleotide hybridizing under stringent conditions with thecomplement of the codin region of SEQ ID NO: 25, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 27, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00220_H05 (SEQ ID NO: 27),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 29, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00222_G03 (SEQ ID NO: 29),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 31 (clone P00223_F07), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 32, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00225_COI (SEQ ID NO: 32),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 34, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00227_D11 (SEQ ID NO: 34),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 36, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00228_F03 (SEQ ID NO: 36),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 38, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00233_H08 (SEQ ID NO: 38),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 40, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00235_G08 (SEQ ID NO: 40),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 42, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00239_C11 (SEQ ID NO: 42),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 44 (clone P00240_B04), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 45, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00240_E05 (SEQ ID NO: 45),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 47 (clone P00241_E12), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 48 (clone P00245_D06), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 49 (clone P00246_D12), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 50, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00247_A04 (SEQ ID NO: 50),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 52, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00248_B04 (SEQ ID NO: 52),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 54, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00249_F09 (SEQ ID NO: 54),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 56, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00258_A10 (SEQ ID NO: 56),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 58, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00262_C10 (SEQ ID NO: 58),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 60 (clone P00263_G06), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 61 (clone P00267_F08) , apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 62, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00269_H08 (SEQ ID NO: 62),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 64 (clone P00312_C04), apolynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 65 (clone P00324_H02), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 66, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00628_H02 (SEQ ID NO: 66),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 68, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00629_C08 (SEQ ID NO: 68),a polynucleotide hybridizing under stringent conditions with thecomplement of the polynucleotide of SEQ ID NO: 70 (clone P00634_G11), apolynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 71, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00641_G11 (SEQ ID NO: 71),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 73, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00648_E12 (SEQ ID NO: 73),a polynucleotide hybridizing under stringent conditions with thecomplement of the coding region of SEQ ID NO: 75, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00697_C03 (SEQ ID NO: 75);(e) a polynucleotide encoding at least about 50 contiguous amino acidsfrom amino acids 1 to 148 of SEQ ID NO: 2, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00184_D11 (SEQ ID NO: 1), a polynucleotideencoding at least about 50 contiguous amino acids from amino acids 1 to193 of SEQ ID NO: 4, wherein said polynucleotide encodes a polypeptidehaving at least one biological activity of the polypeptide encoded byclone P00185_D11 (SEQ ID NO: 3); a polynucleotide encoding at leastabout 50 contiguous amino acids from amino acids 1 to 236 of SEQ ID NO:6, wherein said polynucleotide encodes a polypeptide having at least onebiological activity of the polypeptide encoded by clone P00188_D12 (SEQID NO: 5), a polynucleotide encoding at least about 50 contiguous aminoacids from amino acids 1 to 61 of SEQ ID NO: 8, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 79 of SEQ ID NO: 10, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00194_G01 (SEQ ID NO: 9), a polynucleotideencoding at least about 50 contiguous amino acids from amino acids 1 to92 of SEQ ID NO: 12, wherein said polynucleotide encodes a polypeptidehaving at least one biological activity of the polypeptide encoded byclone P00194_G05 (SEQ ID NO: 11), a polynucleotide encoding at leastabout 50 contiguous amino acids from amino acids 1 to 86 of SEQ ID NO:14, wherein said polynucleotide encodes a polypeptide having at leastone biological activity of the polypeptide encoded by clone P00194_H10(SEQ ID NO: 13), a polynucleotide encoding at least about 50 contiguousamino acids from amino acids 1 to 36 of SEQ ID NO: 16, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00199_D08 (SEQ ID NO: 15),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 83 of SEQ ID NO: 18, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00203_D04 (SEQ ID NO: 17), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 82 of SEQ ID NO: 20, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00203_E06 (SEQ ID NO: 19), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 462 of SEQ ID NO: 22, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00209_F06 (SEQ ID NO: 21), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 170 of SEQ ID NO: 24, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00219_D02 (SEQ ID NO: 23), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids −26 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ ID NO:26), wherein said polynucleotide encodes a polypeptide having at leastone biological activity of the polypeptide encoded by clone P00219_F06(SEQ ID NO: 25), a polynucleotide encoding at least about 50 contiguousamino acids from amino acids 1 to 30 of SEQ ID NO: 28, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00220_H05 (SEQ ID NO: 27),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 39 of SEQ ID NO: 30, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00222_G03 (SEQ ID NO: 29), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 541 of SEQ ID NO: 33, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00225_C01 (SEQ ID NO: 32), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 30 of SEQ ID NO: 35, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00227_D11 (SEQ ID NO: 34), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 100 of SEQ ID NO: 37, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00228_F03 (SEQ ID NO: 36), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 65 of SEQ ID NO: 39, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00233_H08 (SEQ ID NO: 38), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 41 of SEQ ID NO: 39, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00235_G08 (SEQ ID NO: 40), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 46 of SEQ ID NO: 43, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00239_C11 (SEQ ID NO: 42), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 313 of SEQ ID NO: 46, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00240_E05 (SEQ ID NO: 45), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 58 of SEQ ID NO: 51, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00247_A04 (SEQ ID NO: 50), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids −35 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), wherein said polynucleotide encodes a polypeptide having at leastone biological activity of the polypeptide encoded by clone P00248_B04(SEQ ID NO: 52), a polynucleotide encoding at least about 50 contiguousamino acids from amino acids 1 to 58 of SEQ ID NO: 55, wherein saidpolynucleotide encodes a polypeptide having at least one biologicalactivity of the polypeptide encoded by clone P00249_F09 (SEQ ID NO: 54),a polynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 52 of SEQ ID NO: 57, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00258_A10 (SEQ ID NO: 56), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 245 of SEQ ID NO: 59, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00262_C10 (SEQ ID NO: 58), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 142 of SEQ ID NO: 63, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00269_H08 (SEQ ID NO: 62), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 49 of SEQ ID NO: 67, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00628_H02 (SEQ ID NO: 66), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 70 of SEQ ID NO: 69, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00629_C08 (SEQ ID NO: 68), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 113 of SEQ ID NO: 72, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00641_G11 (SEQ ID NO: 71), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 114 of SEQ ID NO: 74, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00648_E12 (SEQ ID NO: 73), apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 97 of SEQ ID NO: 76, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00697_C03 (SEQ ID NO: 75); (f) apolynucleotide encoding at least about 50 contiguous amino acids fromamino acids 1 to 23 of SEQ ID NO: 26, wherein said polynucleotideencodes a polypeptide having at least one biological activity of thepolypeptide encoded by clone P00219_F06 (SEQ ID NO: 25) or amino acids 1to 387 of SEQ ID NO: 53, wherein said polynucleotide encodes apolypeptide having at least one biological activity of the polypeptideencoded by clone P00248_B04 (SEQ ID NO: 52); (g) a polynucleotide of SEQID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32,34, 36, 38, 40, 42, 44, 45, 47, 48, 49, 50, 52, 54, 56, 58, 60, 61, 62,64, 65, 66, 68, 70, 71, 73, and 75; (h) the complement of apolynucleotide of (a)-(g); and (i) an antisense oligonucleotide capableof hybridizing with, and inhibiting the translation of, the mRNA encodedby a gene encoding a polypeptide of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55,57, 59, 63, 67, 69, 72, 74, 76, or another mammalian homologue thereof.2. The polynucleotide of claim 1 encoding a polypeptide comprising aminoacids 1 to 233 of SEQ ID NO: 26, amino acids 1 to 387 of SEQ ID NO: 53.3. The polynucleotide of claim 1 comprising the sequence selected fromthe group consisting of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19,21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40, 42, 44, 45, 47, 48, 49, 50,52, 54, 56, 58, 60, 61, 62, 64, 65, 66, 68, 70, 71, 73, and
 75. 4. Avector comprising and capable of expressing a poly- or oligonucleotideof claim
 1. 5. A recombinant host cell transformed with nucleic acidcomprising a poly- or oligonucleotide of claim
 1. 6. A recombinant hostcell transformed with the vector of claim
 5. 7. A method for producing apolypeptide comprising culturing a recombinant host cell transformedwith nucleic acid comprising any of the polynucleotides of claim1(a)-(g) under conditions such that the polypeptide is expressed, andisolating the polypeptide.
 8. A polypeptide comprising: (a) apolypeptide having at least about 80% identity with amino acids selectedfrom the group consisting of: amino acids 1 to 193 of SEQ ID NO: 4,amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQ ID NO:8, amino acids 1 to 92 of SEQ ID NO:12, amino acids 1 to 86 of SEQ IDNO:14, amino acids 1 to 36 of SEQ ID NO:16, amino acids 1 to 83 of SEQID NO:18, amino acids 1 to 82 of SEQ ID NO:20, amino acids 1 to 462 ofSEQ ID NO:22, amino acids 1 to 170 of SEQ ID NO:24, amino acids 1 to 30of SEQ ID NO:28, amino acids 1 to 30 of SEQ ID NO: 35, amino acids 1 to100 of SEQ ID NO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1to 46 of SEQ ID NO:43, amino acids 1 to 313 of SEQ ID NO:46, amino acids1 to 58 of SEQ ID NO:51, amino acids 1 to 58 of SEQ ID NO:55, aminoacids 1 to 52 of SEQ ID NO:57, amino acids 1 to 245 of SEQ ID NO:59,amino acids 1 to 142 of SEQ ID NO:63, amino acids 1 to 49 of SEQ IDNO:67, amino acids 1 to 70 of SEQ ID NO:69, amino acids 1 to 113 SEQ IDNO:72, and amino acids 1 to 97 of SEQ ID NO:76; or (b) a polypeptideencoded by nucleic acid hybridizing under stringent conditions with thecomplement of the coding region selected from the group consisting of:SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31,32, 34, 36, 38, 40, 42, 44, 45, 47, 48, 49, 50, 52, 54, 56, 58, 60, 61,62, 64, 65, 66, 68, 70, 71, 73, and 75; (c) the polypeptides of (a) and(b) having at least one biological activity of the polypeptide encodedby clones P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12(SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9),P001946_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19),P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ IDNO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01(SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36),P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ IDNO:42), P00240_E05 (SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04(SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56),P00262_C10 (SEQ ID NO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ IDNO:66), P00629_C08 (SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12(SEQ ID NO:73), P00697_C03 (SEQ ID NO:75).
 9. A composition comprising apolypeptide which comprises: (a) a polypeptide having at least about 80%identity with amino acids selected from the group consisting of: aminoacids 1 to 193 of SEQ ID NO: 4, amino acids 1 to 236 of SEQ ID NO:6,amino acids 1 to 61 of SEQ ID NO: 8, amino acids t to 92 of SEQ IDNO:12, amino acids 1 to 86 of SEQ ID NO: 14, amino acids 1 to 36 of SEQID NO: 16, amino acids 1 to 83 of SEQ ID NO218, amino acids 1 to 82 ofSEQ ID NO:20, amino acids 1 to 46 2 of SEQ ID NO:22, amino acids I to170 of SEQ ID NO:24, a mino acids 1 to 30 of SEQ ID NO:28, amino acids 1to 30 of SEQ ID NO: 35, amino acids 1 to 100 of SEQ ID NO:37, aminoacids 1 to 65 of SEQ ID NO:39, amino acids 1 to 46 of SEQ ID NO: 43,amino acids 1 to 313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ IDNO:51, amino acids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQID NO:57, amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 ofSEQ ID NO:37, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70of SEQ ID NO:69, amino acids 1 to 113 of SEQ ID NO:72, and amino acids 1to 97 of SEQ ID NO:76; or (b) a polypeptide encoded by nucleic acidhybridizing under stringent conditions with the complement of the codingregion selected from the group consisting of: SEQ ID NOS: 1, 3, 5, 7, 9,11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40, 42, 44,45, 47, 48, 49, 50, 52, 54, 56, 58, 60, 61, 62, 64, 65, 66, 68, 70, 71,73, and 75; wherein the polypeptides of (a) and (b) have at least onebiological activity of the polypeptide respectively encoded by clonesP00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ IDNO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05(SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15),P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ IDNO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05(SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00225_C01 (SEQ ID NO:32),P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ IDNO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_E05(SEQ ID NO:45), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52),P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ IDNO:58), P00269_H08 (SEQ ID NO:62), P00628_H02 (SEQ ID NO:66), P00629_C08(SEQ ID NO:68), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73),and P00697_C03 (SEQ ID NO:75), in admixture with a carrier.
 10. Thecomposition of claim 9 which is a pharmaceutical composition comprisingan effective amount of said polypeptide in admixture with apharmaceutically acceptable carrier.
 11. An antibody specificallybinding a polypeptide of claim
 8. 12. A composition comprising anantibody of claim 11 in admixture with a carrier.
 13. The composition ofclaim 9 which is a pharmaceutical composition comprising an effectiveamount of said antibody in admixture with a pharmaceutically acceptablecarrier.
 14. A composition comprising an antagonist or an agonist of apolypeptide of claim
 8. 15. The composition of claim 11 which is apharmaceutical composition comprising an effective amount of saidantagonist or said agonist in combination with a pharmaceuticallyacceptable carrier.
 16. A method for the treatment of a cardiac, renalor inflammatory disease, comprising administering to a patient in needan effective amount of a polypeptide of claim 8, or an antagonist oragonist thereof.
 17. A method for the treatment of a cardiac, renal orinflammatory disease, comprising administering to a patient in need aneffective amount of an antibody specifically binding to a polypeptide ofthe present invention.
 18. A method for screening a subject for acardiac, renal or inflammatory disease characterized by the differentialexpression of the polypeptide selected from the group consisting of: SEQID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35,37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, and 76, oran endogenous homologue thereof, comprising the steps of: measuring theexpression in the subject of said polypeptide or said endogenoushomologue; and determining the relative expression of said polypeptideor said endogenous homologue in the subject compared to its expressionin normal subjects, or compared to its expression in the same subject atan earlier stage of development of the cardiac, renal or inflammatorydisease.
 19. The method of claim 15 wherein said subject is human andsaid endogenous homologue is a human homologue of the rat proteinselected from the group consisting of: SEQ ID NOS: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53,55, 57, 59, 63, 67, 69, 72, 74, and
 76. 20. An array comprising one ormore oligonucleotides complementary to reference RNA or DNA encoding aprotein selected from the group consisting of: SEQ ID NOS: 2, 4, 6, 8,10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46,51, 53, 55, 57, 59, 63, 67, 69, 72, 74, and 76, or another mammalian(e.g. human) homologue thereof, where the reference DNA or RNA sequencesare obtained from both a biological sample from a normal subject and abiological sample from a subject exhibiting a cardiac, renal, orinflammatory disease, or from biological samples taken at differentstages of a cardiac, renal, or inflammatory disease.
 21. A method fordetecting cardiac, kidney, or inflammatory disease in a human testpatient comprising the steps of: providing an array of oligonucleotidesat known locations on a substrate, which array comprisesoligonucleotides complementary to reference DNA or RNA sequencesencoding a human homologue of the protein selected from the groupconsisting of: SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24,26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69,72, 74, and 76 where the reference DNA or RNA sequences are obtainedfrom both a biological sample from a normal patient and a biologicalsample from a patient potentially exhibiting cardiac, renal, orinflammnatory disease, or from a test patient exhibiting cardiac, renal,or inflammatory disease, taken at different stages of such disease;exposing the array, under hybridization conditions, to a first sample ofcDNA probes constructed from mRNA obtained from a biological sample froma corresponding biological sample of a normal patient or from a testpatient at a certain stage of the disease; exposing the array, underhybridization conditions, to a second sample of cDNA probes constructedfrom MRNA obtained from a biological sample obtained from the test;quantifying any hybridization between the first sample of cDNA probesand the second sample of cDNA probes with the oligonucleotide probes onthe array; and determining the relative expression of genes encoding thehuman homologue of a protein selected from the group consisting of: SEQID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35,37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, and 76 inthe biological samples from the normal patient and the test patient, orin the biological samples taken from the test patient at differentstages of the disease.
 22. A diagnostic kit for the detection of acardiac, kidney or inflanmmatory disease comprising an array of claim20.
 23. The diagnostic kit of claim 22 further comprising at least oneof the following components: (a) an oligonucleotide probe; (b) a PCRreagent; (c) a detectable label; (d) a biological sample taken from ahuman subject; and (e) an antibody to a polypeptide of any one of thesequences selected from the group consisting of: SEQ ID NOS: 2, 4, 6, 8,10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46,51, 53, 55, 57, 59, 63, 67, 69, 72, 74, 76, and a further mammalianhomologue thereof.
 24. The diagnostic kit of claim 22 wherein saidbiological sample is from blood or a tissue.
 25. The diagnostic kit ofclaim 21 wherein said tissue is a cardiac tissue.
 26. The diagnostic kitof claim 22 wherein said cardiac tissue is a left ventricular tissue.